J Biol Chem, 2000 Mar 17, 275(11), 7497 - 504 Characterization of the linkage between the type III capsular polysaccharide and the bacterial cell wall of group B Streptococcus; Deng L et al.; The capsular polysaccharide of group B Streptococcus is a key virulence factor and an important target for protective immune responses . Until now, the nature of the attachment between the capsular polysaccharide and the bacterial cell has been poorly defined . We isolated insoluble cell wall fragments from lysates of type III group B Streptococcus and showed that the complexes contained both capsular polysaccharide and group B carbohydrate covalently bound to peptidoglycan . Treatment with the endo-N-acetylmuramidase mutanolysin released soluble complexes of capsular polysaccharide linked to group B carbohydrate by peptidoglycan fragments . Capsular polysaccharide could be enzymatically cleaved from group B carbohydrate by treatment of the soluble complexes with beta-N-acetylglucosaminidase, which catalyzes hydrolysis of the beta-D-GlcNAc(1-->4)beta-D-MurNAc subunit produced by mutanolysin digestion of peptidoglycan . Evidence from gas chromatography/mass spectrometry and (31)P NMR analysis of the separated polysaccharides supports a model of the group B Streptococcus cell surface in which the group B carbohydrate and the capsular polysaccharide are independently linked to the glycan backbone of cell wall peptidoglycan; group B carbohydrate is linked to N-acetylmuramic acid, and capsular polysaccharide is linked via a phosphodiester bond and an oligosaccharide linker to N-acetylglucosamine. Vet Pathol, 2000 Mar, 37(2), 143 - 52 Pathogenesis of porcine reproductive and respiratory syndrome virus-induced increase in susceptibility to Streptococcus suis infection; Thanawongnuwech R et al.; Eighty 3-week-old crossbred pigs were randomly assigned to six groups (13-14 pigs/group) . Group 1 pigs served as uninoculated controls, group 2 pigs were inoculated intranasally (i.n.) with Streptococcus suis serotype 2, group 3 pigs were inoculated i.n . with a modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine, group 4 pigs were inoculated i.n . with the same vaccine and with S . suis, group 5 pigs were inoculated i.n . with VR-2385 (a high-virulence strain of PRRSV), and group 6 pigs were inoculated i.n . with VR-2385 and S . suis . Pigs exposed to both PRRSV and S . suis were inoculated with PRRSV 7 days prior to S . suis inoculation . The pigs were 26 days old when inoculated with S . suis . Respiratory disease was significantly more severe in groups 5 and 6 . Mortality rate was the highest in group 6 (87.5%) . This rate was significantly higher than that observed in all other groups except group 4 (37.5%) . The mortality rate in group 2, inoculated with S . suis alone, was 14.3% . No pigs from groups 1, 3, or 5 died prior to the scheduled necropsies at 10 and 28 days postinoculation with PRRSV (DPI) . To study the effect of PRRSV and/or S . suis on pulmonary clearance by pulmonary intravascular macrophages, six pigs from each group were intravenously infused with 3% copper phthalocyanine tetrasulfonic acid in saline prior to necropsy at 10 DPI . Mean copper levels in the lungs of pigs in groups 2, 5, and 6 were significantly lower than those in control pigs . The mean percentage of lung tissue grossly affected by pneumonia at 10 DPI was 0%, 1%, 0%, 3%, 64%, and 62% for groups 1-6, respectively . Both gross and microscopic interstitial pneumonia lesions were significantly more severe in the VR2385-inoculated groups (5 and 6) . PRRSV was isolated from bronchoalveolar lavage fluid collected at necropsy from 100% of the pigs in groups 5 and 6, 71.4% of pigs in group 4, 38.5% of pigs in group 3, and none of the pigs in groups 1 or 2 . Streptococcus suis serotype 2 was cultured from the internal tissues of 7.7%, 28.6%, and 78.6% of the pigs in groups 2, 4, and 6, respectively . Streptococcus suis serotype 2 was isolated from whole blood at necropsy from 7.7%, 35.7%, and 78.6% of pigs in groups 2, 4, and 6, respectively . Significantly more pigs in group 6 had S . suis isolated from whole blood and internal tissues . In summary, both high-virulence PRRSV and S . suis decreased copper clearance, and the incidence of isolation of S . suis and PRRSV was higher in dually inoculated pigs . PRRSV-induced suppression of pulmonary intravascular macrophage function may in part explain PRRSV-associated increased susceptibility to S . suis infection. Mol Microbiol, 2000 Mar, 35(5), 1089 - 98 Regulation of competence for genetic transformation in Streptococcus pneumoniae: expression of dpnA, a late competence gene encoding a DNA methyltransferase of the DpnII restriction system; Lacks SA et al.; The chromosomal DpnII gene cassette of Streptococcus pneumoniae encodes two methyltransferases and an endonuclease . One methyltransferase acts on double-stranded and the other on single-stranded DNA . Two mRNAs are transcribed from the cassette . One, a SigA promoter transcript, includes all three genes; the other includes a truncated form of the second methyltransferase gene (dpnA) and the endonuclease gene . The truncated dpnA, which is translated from the second start codon in the full gene, was shown to produce active enzyme . A promoter reporter plasmid for S . pneumoniae was devised to characterize the promoter for the second mRNA . This transcript was found to depend on a promoter that responded to the induction of competence for genetic transformation . The promoter contains the combox sequence recognized by a SigH-containing RNA polymerase . As part of the competence regulon, the dpnA gene makes a product able to methylate incoming plasmid strands to protect them from the endonuclease and allow plasmid establishment . Its function differs from most genes in the regulon, which are involved in DNA uptake . Comparison of R6 and Rx strains of S . pneumoniae showed the temperature dependence of transformation in R6 to result from temperature sensitivity of the uptake apparatus and not the development of competence. Am J Respir Crit Care Med, 2000 Mar, 161(3 Pt 1), 753 - 62 Treatment of gram-positive nosocomial pneumonia . Prospective randomized comparison of quinupristin/dalfopristin versus vancomycin . Nosocomial Pneumonia Group; Fagon J et al.; Nosocomial pneumonia is a frequent complication in hospitalized patients . Gram-positive pathogens, particularly Staphylococcus aureus, are responsible for the increasing frequency of nosocomial pneumonia . To evaluate the efficacy and safety of intravenous quinupristin/dalfopristin (Synercid) in the treatment of nosocomial pneumonia caused by gram-positive pathogens we conducted a prospective, randomized, open-label, international, multicenter, comparative clinical trial . Two hundred ninety-eight patients with nosocomial pneumonia were enrolled in 74 active centers in five countries: a subgroup of 171 (87 quinupristin/dalfopristin-treated and 84 vancomycin-treated patients) were evaluable for the major efficacy end points . One hundred fifty received 7.5 mg/kg of quinupristin/dalfopristin every 8 h; 148 patients received 1 g of vancomycin every 12 h . Aztreonam at a dose of 2 g every 8 h could be administered in both groups for coverage of gram-negative organisms, and tobramycin was added for coverage against Pseudomonas aeruginosa . The primary efficacy end point was the clinical response between the seventh and the thirteenth day after the end of treatment in clinically evaluable patients with documented causative pathogen(s) at baseline (bacteriologically evaluable population) . Therapy was clinically successful (cure or improvement) in 49 (56.3%) of patients receiving quinupristin/dalfopristin and 49 (58.3%) patients receiving vancomycin (difference, -2.0% {95% CI, -16.8% to 12.8%}) in the bacteriologically evaluable population . Equivalent clinical success rates were also observed in the all-treated population (n = 298), and in the bacteriologically evaluable patients intubated in baseline (39/72 {54%} compared with 36/67 {54%}) . The by-pathogen bacteriologic response was similar in both treatment groups, with equivalent clinical success rates for Streptococcus pneumoniae, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus . Adverse events (venous and nonvenous) were equally distributed between groups; 15.3% of quinupristin/dalfopristin patients and 9.5% of vancomycin patients discontinued therapy because of an adverse clinical event . In this study quinupristin/dalfopristin was shown to be equivalent to vancomycin in the treatment of nosocomial pneumonia caused by gram-positive pathogens . Quinupristin/dalfopristin merits further evaluation for the treatment of nosocomial pneumonia caused by methicillin-resistant S . aureus. Anim Reprod Sci, 2000 Mar 15, 58(3-4), 241 - 51 Endocrine profiles of dairy cows following experimentally induced clinical mastitis during early lactation; Hockett ME et al.; Concentrations of LH, cortisol, estradiol-17beta (E(2)), prolactin and 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM) were determined in cows with experimentally induced clinical mastitis during early lactation . Cows free of intramammary infection (IMI) and in the luteal phase of the estrous cycle were balanced by lactation number and days in milk and assigned to either control (n=5) or treatment (n=5) groups . Treated cows were infected experimentally (day 0), in two mammary quarters, with Streptococcus uberis and developed clinical mastitis within 60 h after inoculation as evidenced by increased mastitis scores, elevated rectal temperatures, mammary swelling and isolation of S . uberis pathogen . Four days following bacterial challenge, blood samples were collected every 20 min for 8 h for determination of PGFM and LH following administration of oxytocin and GnRH, respectively . Blood samples were also collected on days 0, 4 and 7 of the experiment to determine concentrations of E(2), prolactin and cortisol . Four days after bacterial challenge, concentrations of cortisol were higher (P=0.04) in experimentally infected cows than controls . Experimentally challenged cows had increased (P=0.02) concentrations of cortisol on days 4 and 7 compared with day 0 . Control cows had no significant increase in blood cortisol during the experimental period . Baseline concentrations of PGFM did not differ between groups; however, peak concentrations of PGFM following oxytocin challenge were elevated (P=0.006) in cows with clinical mastitis compared with control animals . Prolactin, E(2) and LH did not differ between cows with clinical mastitis or controls . Experimentally induced mastitis during early lactation elevated concentrations of cortisol during the luteal phase of the estrous cycle . Furthermore, mastitic cows demonstrated an increased PGFM response following oxytocin administration . Altered reproductive efficiency in cows with clinical mastitis caused by Gram-positive pathogens may be the result of increased uterine sensitivity to prostaglandin F(2alpha) (PGF(2alpha)). Crit Care Med, 2000 Feb, 28(2), 377 - 82 Nosocomial endocarditis in the intensive care unit: an analysis of 22 cases; Gouello JP et al.; OBJECTIVES: To review the intensive care unit experience of patients with admitted or acquired nosocomial endocarditis (NE) defined according to the Duke criteria . DESIGN: Prospective, cohort study . SETTING: University teaching hospital . PATIENTS: We reviewed the records of 22 patients documented with NE during a 6-yr period from 1992 to 1997 . INTERVENTIONS: None . MEASUREMENTS AND MAIN RESULTS: Twenty-two patients (9 women/13 men) aged 38-83 yrs (mean 65+/-9 yrs) had a NE (prevalence of 5 per 1,000 admissions) . For six patients, NE was the reason for the admission to the intensive care unit . For 17 patients, the time elapsed between admission and diagnosis of NE was 39+/-25 days . Sixteen patients were predisposed to infection and seven had underlying heart conditions that put them at risk for acute endocarditis: three prosthetic valves, two valvular diseases, and two cardiac pacemakers . In 21 cases (one unknown portal of entry), NE was the consequence of bacteremia related to a medical or surgical procedure: 11 intravascular devices, eight surgical wounds, one tracheal procedure, and one leg ulceration . The bacteriologic agents detected in blood cultures were: staphylococci (n = 17), Streptococcus (n = 2), Pseudomonas aeruginosa (n = 2), and Candida (n = 2) . Fourteen patients underwent echocardiography according to cardiac signs (cardiac failure, new cardiac murmur, or embolic event) . For the eight remainders, echocardiography was performed systematically because of fever and positive blood cultures . The lesions detected by 21 transthoracic and 17 transesophageal echocardiographs were the following: vegetations (n = 19), myocardial abscesses (n = 5), and valvular perforation (n = 1) . On 16 surgical indications, only five patients underwent surgery because the others were in too poor of a condition . The overall mortality was 68% (n = 15) and was directly associated with NE in 36% of cases (n = 8) . Seven patients (28%) were discharged 34 days after the diagnosis of endocarditis . CONCLUSIONS: NE is a frequent nosocomial infection that occurs late during hospitalization . Persistent fever with positive blood cultures is sufficient symptomology to promptly perform an echocardiogram . The poor prognosis is related to the poor condition of those patients who cannot be referred for surgical treatment. J Med Microbiol, 2000 Mar, 49(3), 245 - 52 Epithelial cell response to challenge of bacterial lipoteichoic acids and lipopolysaccharides in vitro; Pollanen MT et al.; Accumulating dental plaque at the gingival margin contains lipoteichoic acids (LTAs) from the cell walls of gram-positive bacteria . In subgingival plaque associated with periodontal disease the amount of lipopolysaccharides (LPSs) from gram-negative bacteria increases . As the gingival junctional epithelium (JE) is an important structural and functional tissue, participating in the first line defence against apical advancement of dental plaque, this study examined the direct effects of LTAs (from Streptococcus mutans and S . sanguis) and LPSs (from Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola and Escherichia coli) on two epithelial cell lines (HaCaT and ERM) and a culture model for human JE . The cells were exposed to the LTAs or LPSs (10-50 microg/ml) for variable periods of time . None of the bacterial surface components had any effect on primary adhesion or on the epithelial attachment of the JE cultures . However, cell growth and mitotic activity were consistently reduced in all cultures treated with LTAs . In contrast, LPSs showed only slight or no effects on cell growth and mitotic activity depending on the epithelial cells used . This suggests that LPSs, despite their established role as modulators of inflammation, do not have direct harmful effects - at the concentrations found in dental plaque and gingival crevicular fluid - which would explain the mechanism of epithelial degeneration and detachment from the tooth surface . However, the LTAs appear to inhibit the renewal of epithelium and may thus contribute to degeneration of coronal JE and subgingival colonisation by periodontal pathogens. N Engl J Med, 2000 Mar 9, 342(10), 681 - 9 Cigarette smoking and invasive pneumococcal disease . Active Bacterial Core Surveillance Team; Nuorti JP et al.; BACKGROUND: Approximately half of otherwise healthy adults with invasive pneumococcal disease are cigarette smokers . We conducted a population-based case-control study to assess the importance of cigarette smoking and other factors as risk factors for pneumococcal infections . METHODS: We identified immunocompetent patients who were 18 to 64 years old and who had invasive pneumococcal disease (as defined by the isolation of Streptococcus pneumoniae from a normally sterile site) by active surveillance of laboratories in metropolitan Atlanta, Baltimore, and Toronto . Telephone interviews were conducted with 228 patients and 301 control subjects who were reached by random-digit dialing . RESULTS: Fifty-eight percent of the patients and 24 percent of the control subjects were current smokers . Invasive pneumococcal disease was associated with cigarette smoking (odds ratio, 4.1; 95 percent confidence interval, 2.4 to 7.3) and with passive smoking among nonsmokers (odds ratio, 2.5; 95 percent confidence interval, 1.2 to 5.1) after adjustment by logistic-regression analysis for age, study site, and independent risk factors such as male sex, black race, chronic illness, low level of education, and living with young children who were in day care . There were dose-response relations for the current number of cigarettes smoked per day, pack-years of smoking, and time since quitting . The adjusted population attributable risk was 51 percent for cigarette smoking, 17 percent for passive smoking, and 14 percent for chronic illness . CONCLUSIONS: Cigarette smoking is the strongest independent risk factor for invasive pneumococcal disease among immunocompetent, nonelderly adults . Because of the high prevalence of smoking and the large population attributable risk, programs to reduce both smoking and exposure to environmental tobacco smoke have the potential to reduce the incidence of pneumococcal disease. Vet Rec, 2000 Feb 5, 146(6), 159 - 61 Detection of DNA restriction fragment polymorphisms in Streptococcus equi; Takai S et al.; Large-restriction-fragment (LRF) polymorphisms in Streptococcus equi (S equi subspecies equi) were studied by pulsed-field gel electrophoresis . Five or six chromosomal fragments of between 194 and 915 kb were separated by digestion with the restriction endonuclease Notl . All 20 isolates of S equi, including 12 from independent Japanese outbreaks, four from independent American outbreaks, two from a single Irish outbreak, us vaccine strain F43, and type strain NCTC 9682 were successfully typed . Seven distinctive, reproducible and stable types were identified . The 12 Japanese isolates collected between 1992 and 1998 were of LRF type II suggesting that they were derived from the same source . The remaining eight isolates were of six types . The results indicate that LRF typing should be a useful technique for investigating the source and transmission of S equi. J Community Health, 2000 Feb, 25(1), 23 - 33 Predicting cost-benefits before programs are started: looking at conjugate vaccine for invasive pneumococcal infections; Hueston WJ et al.; This analysis uses existing data to examine how an analysis to predict the net financial impact for an emerging medical program, namely a conjugate vaccine against Streptococcus pneumoniae, and to identify which key variables will have the greatest impact on the program's costs and benefits . Using data available on the prevalence and case fatality rates for invasive diseases caused by S pneumoniae, we examined the theoretical economic impact of vaccinating all newborns versus not vaccinating . Effectiveness estimates for conjugated pneumococcal vaccines and disease incidence and fatality rates were obtained from published sources . Because of scanty or inconclusive data for otitis media and pneumonia, the analysis was limited to cases of meningitis and bacteremia due to S pneumoniae . Based on these two diseases alone, immunization with conjugate pneumococcal vaccine could save an estimated 222 lives per million children vaccinated per year . Analysis of direct costs (projected immunization costs minus savings from reduced illness) show that a pneumococcal vaccine program will result in net direct costs between $0.08 and $2.42 per child . When indirect costs are included in the analysis, the vaccine is cost savings for all cases except when the two year incidence of disease and death rates are lowest and the cost of the vaccine series is $150 . Further research should focus on these key issues as the vaccine is introduced into use, as expected in the next few years. Zentralbl Bakteriol, 2000 Jan, 289(8), 835 - 43 Complementary characterization of a hyaluronic acid splitting enzyme from Streptococcus agalactiae; Rodig H et al.; A hyaluronic acid splitting enzyme of Streptococcus agalactiae was characterized by splitting mechanism, Michaelis-constant and inhibition type for sulfated hyaluronic acid: The enzyme splits hyaluronic acid as a hyaluronate lyase {EC 4.2.2.1} . The Km = 8 x 10(-4) mg ml-1 was determined with the influence of substrate inhibition constant Kiu = 2 x 10(-6) mg ml-1 . Sulfated hyaluronic acid inhibits the enzyme in a partially non-competitive way . The inhibition constant is Ki = 5.47 x 10(-4) mg ml-1 . The GBS-hyaluronate lyase cleaves hyaluronic acid as an endoglycosidase . The work is related with the intention to establish a hyaluronate lyase of microbial origin as a therapeutical enzyme replacing bovine hyaluronidase. Diagn Microbiol Infect Dis, 2000 Feb, 36(2), 81 - 4 Streptococcus pneumoniae from ophthalmic infections: serotype distribution and penicillin susceptibility; Mathews MS et al.; Streptococcus pneumoniae is one of the pathogens causing infection of the conjunctiva and the uveal tract . The present study began with the observation of two ophthalmic S . pneumoniae isolates showing intermediate resistance to penicillin . Among the 25 isolates of S . pneumoniae from 617 ophthalmic specimens, during the period of 14 months, four were found to exhibit an intermediate resistance to penicillin . Minimum Inhibitory Concentration values ranging from 0.125 microg/mL to 0.25 microg/mL was observed . No multidrug resistant strains were isolated . Serogrouping/typing of the S . pneumoniae revealed the following serogroups/types; 6A (n = 3), 6B (n = 2), 22 (n = 3), 14 (n = 3), 23A (n = 2), and 1 each of 23B, 19A, 7B, 32, 9, 42, 21, 39, 10, 3, and 34 . One strain showed cross reaction in pool 29, 35, and 47 . These findings represent the first such observation of ophthalmic isolates from India. Mol Gen Mikrobiol Virusol, 2000, (1), 23 - 7 {New rare cutting restriction endonuclease SmII from Streptococcus milleri recognises 5'-ATTTAAAT-3'}; Dedkov VS et al.; New restriction endonuclease (restrictase) Smil of type II was detected in the bacterial strain Streptococcus milleri . Cellular lysate enzyme cut T7 and adenovirus-2 DNAs at site 5'-ATTT decreases AAAT-3' but not lambda DNA which does not contain this sequence . Intense aeration inhibited the growth of S . milleri . The content of restrictase in the cells was the greatest during the logarithmic growth phase . A total of 20,000 units of Smil were isolated from 4 g of cells by cellular extract fractionation with ammonium sulfate and subsequent chromatography on columns with Bio Gel A 0.5 m, heparin agarose, and phosphocellulose . Purified enzyme cut the synthetic oligonucleotide duplex in the center of the recognized site 5'-ATTT decreases AAAT-3' . Smil restrictase is a true isoschisomer of rare-cutting Swal enzyme . Smil belongs to a small group of enzymes which recognize octanucleotide sites and can be used for large-block fragmentation of DNA . Comparison of specificities of rare-cutting and other restrictases suggests that the enzymes recognizing octanucleotides can evolutionally originate from enzymes recognizing both hexanucleotides and tetranucleotides. J Antimicrob Chemother, 2000 Mar, 45(3), 379 - 82 Therapeutic efficacy of J-111,225, a novel trans-3,5-disubstituted pyrrolidinylthio-1beta-methylcarbapenem, against experimental murine systemic infections; Shibata K et al.; In a murine model of systemic infection with methicillin-resistant Staphylococcus aureus (MRSA), J-111,225 showed an ED(50) value of 5 . 83 mg/kg, which was comparable to vancomycin (ED(50) 4.84 mg/kg), whereas imipenem failed to cure infected mice (ED(50) >100 mg/kg) . Against a mixed infection caused by MRSA and Pseudomonas aeruginosa, monotherapy with J-111,225 showed an ED(50) value of 7.23 mg/kg, whereas combined treatment with vancomycin plus imipenem (1:1) had an ED(50) of 20.86 mg/kg . J-111,225 showed good therapeutic efficacy against methicillin-susceptible S . aureus, penicillin-resistant Streptococcus pneumoniae, Escherichia coli, Klebsiella pneumoniae and P . aeruginosa . The unusually broad spectrum suggests that monotherapy with this novel carbapenem may be suitable for polymicrobial infections associated with MRSA. J Antimicrob Chemother, 2000 Mar, 45(3), 315 - 20 Evaluation of combined ceftriaxone and dexamethasone therapy in experimental cephalosporin-resistant pneumococcal meningitis; Cabellos C et al.; The treatment of meningitis caused by strains of Streptococcus pneumoniae with decreased susceptibility to third-generation cephalosporins is an increasingly frequent and difficult problem . In this study a rabbit model of meningitis was used to determine the efficacy of ceftriaxone at different dosages, and to establish the effect of the addition of dexamethasone to the chemotherapeutic regimen . Groups of eight rabbits were inoculated with 10(6) cfu/mL of a cephalosporin- resistant strain of S . pneumoniae (MIC of cefotaxime/ceftriaxone 2 mg/L) . Eighteen hours after inoculation, ceftriaxone (50 or 100 mg/kg/day) with or without dexamethasone (0 . 25 mg/kg/ day) was administered for a period of 48 h . The ceftriaxone dose of 50 mg/kg/day was not fully effective in this model (therapeutic failure rate 28%) . With a dose of 100 mg/kg/day there were no therapeutic failures and all CSF cultures were below the level of detection at 48 h . CSF ceftriaxone concentrations, area under the time-concentration curve and time above the MIC were not significantly different with or without dexamethasone . However, concomitant use of dexamethasone resulted in higher CSF bacterial counts and a higher number of therapeutic failures (57% with the 50 mg/kg/day dose and 28% with the 100 mg/kg/day dose) . Increasing doses of ceftriaxone might be an effective mode of therapy for meningitis caused by S . pneumoniae with MIC </= 2 mg/L . However, in contrast to cephalosporin-sensitive cases, in cases caused by ceftriaxone-resistant strains, concomitant use of dexamethasone was associated with a higher failure rate even when a higher dosage of ceftriaxone was used. J Antimicrob Chemother, 2000 Mar, 45(3), 311 - 4 Therapeutic effects of parenteral beta-lactam antibiotics on experimental otitis media caused by penicillin-resistant Streptococcus pneumoniae in guinea-pigs; Hori R et al.; The therapeutic effects of parenteral beta-lactam antibiotics were evaluated in experimental acute otitis media caused by penicillin-resistant Streptococcus pneumoniae (PRSP) in guinea-pigs . Cefotaxime, ceftriaxone and piperacillin significantly reduced viable cell counts of PRSP in the middle ear at a dose of 50 mg/kg bd for 3 days (P < 0.01 compared with control) . The therapeutic effects of cefotaxime, ceftriaxone and piperacillin were superior to those of cefotiam and ceftazidime . These therapeutic effects reflected both in vitro activity and pharmacokinetic properties of the drugs. Clin Diagn Lab Immunol, 2000 Mar, 7(2), 314 - 7 Comparison of polymorphonuclear cells from healthy donors and differentiated HL-60 cells as phagocytes in an opsonophagocytic assay using antigen-coated fluorescent beads; Guy B et al.; Polymorphonuclear cells (PMNs) from healthy donors and differentiated HL-60 cells were compared in an opsonophagocytic assay using fluorescent latex beads coated with Streptococcus pneumoniae polysaccharide conjugates . Serum-specific phagocytosis was efficiently mediated by both sources of cells, as measured by flow cytometry, but the mean number of beads ingested per cell was three- to fivefold higher when PMNs were used than when HL-60 cells were used . Nevertheless, differentiated HL-60 cells could be a convenient and standardized source of cells to evaluate the functionality of specific antibodies to vaccine candidates as a coating on fluorescent beads. Microb Pathog, 2000 Mar, 28(3), 135 - 44 Sequence and characterization of the glyceraldehyde-3-phosphate dehydrogenase of Mycobacterium avium: correlation with an epidermal growth factor binding protein; Parker AE et al.; Mycobacterium avium is a common pathogen in AIDS patients . The extracellular environment within the granuloma shown to support mycobacterial growth is in the caseous fluid . Previous work demonstrated that the presence of human epidermal growth factor (EGF), which is found in the tissue of chronic granulomous lesions, increases the growth rate of M . avium and Mycobacterium tuberculosis . Previously, a protein capable of binding recombinant human EGF (rEGF) in a western blot was identified with homology to glyceraldehyde-3-phosphate dehydrogenase (GAP) in both M . avium and M . tuberculosis but not Mycobacterium smegmatis . Surface GAPs have been identified in group A Streptococcus, enteropathogenic Escherichia coli, Candida albicans and Schistosoma mansoni . We have cloned the gap gene of M . avium . M . avium GAP has high homology with M . tuberculosis GAP . The protein was also expressed in M . smegmatis, conveying the ability to bind rEGF, but no growth increase was observed in 7H9 broth in the presence of rEGF up to 500 ng/ml . Only one copy of the GAP gene was identified in M . avium These results contribute to the understanding of M . avium pathogenesis by characterizing its interaction with a host protein present in the site of infection . Anaesth Intensive Care, 2000 Feb, 28(1), 87 - 90 Streptoccocus pyogenes: a forgotten cause of severe community-acquired pneumonia; Birch C et al.; We report a case of severe community-acquired pneumonia caused by Streptococcus pyogenes (Lancefield Group A streptoccocus) that was complicated by a streptococcal toxic shock syndrome . Although this micro-organism is an uncommon cause of community-acquired pneumonia, previously well individuals may be infected and the clinical course may be fulminant . A household contact was the likely point of infection . Invasive group A streptococcal disease continues to remain an important cause of morbidity and mortality in the community and therefore will continue to be encountered by intensive care physicians . Treatment of Group A streptococcal infection remains penicillin; however, clindamycin should be added in severe infection. Vaccine, 2000 Mar 17, 18(18), 1873 - 85 Preparation of pneumococcal capsular polysaccharide-protein conjugate vaccines utilizing new fragmentation and conjugation technologies; Pawlowski A et al.; There is a global urgent need for a new efficient and inexpensive vaccine to combat pneumococcal disease, which should also be affordable in developing countries . In view of this need a simple low-cost technique to prepare such a vaccine was developed . The preparation of serotype 14 and 23F pneumococcal capsular polysaccharide (PnPS)-protein conjugates to be included in a forthcoming multivalent PnPS conjugate vaccine is described . Commercial lots of PnPSs produced according to Good Manufacturing Practice from Streptococcus pneumoniae serotype 14 (PS14) and 23F (PS23F) were partially depolymerized by sonication or irradiation in an electron beam accelerator . The PnPS fragments were conjugated to tetanus toxoid (TT) using a recently developed conjugation chemistry . The application of these new simple, efficient and inexpensive fragmentation and conjugation technologies allowed the synthesis of several PnPS-protein conjugates containing PnPS fragments of preselected sizes and differing in the degree of substitution . The PS14TT and PS23FTT conjugate vaccine candidates were characterized chemically and their immunogenicity was evaluated in rabbits and mice . All PnPS conjugate vaccines, unlike the corresponding plain polysaccharides, produced high IgG titres in both animal species . The PS14TT conjugates tended to be more immunogenic than the PS23FTT conjugates . The immune response to the PS14TT conjugates, but not to the PS23FTT conjugates, was related to the size of the conjugated polysaccharide hapten . Both types of conjugates elicited strong booster effects upon secondary immunizations, resulting in high IgG1, IgG2a and IgG2b titres. Vaccine, 2000 Mar 6, 18(17), 1811 - 21 Purification and characterization of Streptococcus pneumoniae palmitoylated pneumococcal surface adhesin A expressed in Escherichia coli; De BK et al.; All Streptococcus pneumoniae isolates tested to date express a species-common lipoprotein designated as pneumococcal surface adhesin A (PsaA) . This protein is cell-associated, hydrophobic, immunogenic, and genetically conserved . It is currently under investigation as a potential component in third-generation pneumococcal vaccine formulations . To overcome the problem of low-level expression of native hydrophobic PsaA in S . pneumoniae, and also of the recombinant PsaA (rPsaA) in Escherichia coli, we generated a stable E . coli construct expressing functional palmitoylated rPsaA ( approximately 10 mg/l of fermentation culture) using Borrelia burgdorferi outer surface protein A (OspA, a hydrophobic lipoprotein) signal peptide . By Western blot analysis, the chimeric rPsaA ( approximately 34 kDa) was detected in the cell lysate using anti-PsaA antibodies . It was partially purified by extracting the cell pellet with PBS/Triton X(R)-114 buffers, followed by anion exchange filter chromatography . A trypsin digestion profile of rPsaA closely resembled that of the native protein, as revealed by SDS-PAGE/silver staining . Lipidation of rPsaA was confirmed by labeling recombinant E . coli cells with {(3)H} palmitic acid and analyzing the labeled E . coli cells by Western blotting coupled with autoradiography . Further, analysis of purified rPsaA by mass spectrometry (MALDI-TOF) revealed a heterogenous spectrum with a major peak (M+H)(+1) of mass 33,384 Da (theoretical mass of palmitoylated rPsaA=33,361 Da) . Purified rPsaA was immunogenic in CBA/NCAHN-XID female mice following intranasal immunization with or without adjuvant, as determined by measurement of anti-PsaA serum IgG levels . These anti-PsaA antibodies reacted with both native and rPsaA polypeptides . Our data strongly suggest that E . coli-expressed rPsaA is palmitoylated and closely resembles the native protein in structure and immunogenicity . It was also observed to elicit measurable protection against nasopharyngeal carriage with S . pneumoniae. Pediatrics, 2000 Mar, 105(3 Pt 1), 502 - 9 Reevaluation of outpatients with Streptococcus pneumoniae bacteremia; Bachur R et al.; BACKGROUND: The reevaluation process for outpatients recalled for Streptococcus pneumoniae bacteremia has not been standardized . Children who return ill or with new serious focal infections require admission and parenteral antibiotic therapy . Limited data exist to guide the follow-up management of those patients identified as having occult pneumococcal bacteremia . OBJECTIVES: Characterize the outcomes of outpatients with pneumococcal bacteremia based on their evaluation at follow-up . For patients who are well-appearing without serious focal infection, propose a management scheme for reevaluation . METHODS: Retrospective review of outpatients with pneumococcal bacteremia . Patients with immunocompromise, those identified with focal bacterial infection at the initial visit, or those admitted at the initial visit were excluded . Data were collected from the initial visit (when blood culture drawn) and follow-up visit with regard to clinical parameters, laboratory data, diagnoses, and any antibiotic treatment . Decision tree analysis was used to generate a model to predict children at high risk for persistent bacteremia (PB) . RESULTS: A total of 548 episodes of pneumococcal bacteremia were studied . Seventy-three children received no antibiotic, 239 oral antibiotic, and 236 parenteral antibiotic at the initial visit . Median age, temperature, and white blood cell (WBC) count were 13.5 months, 40.0 degrees C, and 20 400/mm(3) . Forty-one patients had PB or new focal infections (15 with PB alone, 4 had focal infection and PB) . Eight patients had meningitis at follow-up . Ninety-two percent returned because of notification of the positive blood culture result . A repeat blood culture was obtained in 92%, 23% had a lumbar puncture, 33% had a chest radiograph, and 12% were admitted . PB was associated with the antibiotic treatment group, elevation of temperature, and WBC count at follow-up . A simple management scheme using 2 sequential decision nodes of antibiotic treatment (none vs any) and then temperature at follow-up (>38.8 degrees C) would have predicted 16/19 patients with PB (sensitivity =.84 and specificity =.86) . CONCLUSIONS: All patients with pneumococcal bacteremia need prompt reevaluation . For well-appearing patients without new focal infection, the utility of diagnostic testing (specifically repeat blood cultures) and the need for admission may be determined by the use of antibiotics at the initial evaluation and the presence of fever at follow-up . The majority of patients can be managed as outpatients entirely . Patients who did not receive antibiotics at the initial evaluation and those treated with oral antibiotics but remain febrile are at the highest risk for persistent bacteremia. Infection, 2000 Jan-Feb, 28(1), 58 - 9 Decreased susceptibility to extended-spectrum cephalosporins of a penicillin-susceptible Streptococcus pneumoniae in meningitis; De Champs C et al.; A 69-year-old woman was admitted to the hospital with meningitis due to Streptococcus pneumoniae . The strain was susceptible to penicillin but intermediate to cefotaxime . In Europe the decrease of susceptibility generally pertains more to penicillin than to cefotaxime . Such a strain is perhaps a forewarning of the existence of high-level cephalosporin-resistant strains . Despite the possible detection of the resistance by oxacillin disk, it underlines the need to determine the MICs of different beta-lactams without delay and to choose the most efficient treatment. Infection, 2000 Jan-Feb, 28(1), 51 - 2 Double prosthetic valve endocarditis caused by Streptococcus pneumoniae; Claes K et al.; Infective endocarditis (IE) caused by Streptococcus pneumoniae is a rare disease . Only eight cases of pneumococcal prosthetic valve endocarditis have been described in the literature . In this report we describe the first case of pneumococcal endocarditis involving two prosthetic heart valves . The patient had pneumonia as the probable portal of entry but no predisposing conditions for invasive pneumococcal disease . Our case also illustrates the importance of transesophageal echocardiography (TEE) for the early diagnosis of IE and a timely decision for cardiac surgery. Infection, 2000 Jan-Feb, 28(1), 13 - 20 Release of teichoic and lipoteichoic acids from 30 different strains of Streptococcus pneumoniae during exposure to ceftriaxone, meropenem, quinupristin/dalfopristin, rifampicin and trovafloxacin; Heer C et al.; The release of teichoic acids (TA) and lipoteichoic acids (LTA) from 30 different strains of Streptococcus pneumoniae during exposure to ceftriaxone, meropenem, quinupristin/dalfopristin, rifampicin and trovafloxacin at concentrations of 10 micrograms/ml and of the respective MIC was determined by an enzyme immunoassay . At 10 micrograms/ml the most rapid and intense release was detected during treatment with the beta-lactam antibiotics ceftriaxone and meropenem, the lowest release was seen with rifampicin and quinupristin/dalfopristin . Trovafloxacin delayed the release of TA/LTA . The maximum concentrations of TA/LTA, however, during trovafloxacin treatment were almost as high as those during exposure to ceftriaxone and meropenem . During exposure to the MIC, ceftriaxone, meropenem, rifampicin and trovafloxacin released significantly higher amounts of TA/LTA than during exposure to 10 micrograms/ml (p < 0.01) . Only quinupristin/dalfopristin released small amounts of TA/LTA at the low and high concentration . In conclusion, at high concentrations antibiotics that do not affect the bacterial cell wall released less pro-inflammatory compounds from S . pneumoniae than ceftriaxone and meropenem . This may be of value in the treatment of meningitis and sepsis. Anticancer Res, 1999 Nov-Dec, 19(6C), 5511 - 4 Inhalation therapy using a streptococcal preparation (OK-432) against bronchioloalveolar carcinoma of the lung; Yano T et al.; OBJECTIVE: In order to inhibit the occurrence of airborne metastasis of bronchioloalveolar cell carcinoma (BAC), we tried to activate alveolar macrophages by the inhalation of aerosolized OK-432, which is a heat and penicillin-treated lyophilized preparation of the Su strain of Streptococcus pyogenes . METHODS AND RESULTS: Alveolar macrophages were obtained from resected specimens of lung cancer patients and cultured for 24 h in the presence of various concentrations of OK-432 (0.001-1 KE/ml) . The cytotoxic activity against a lung cancer cell line was augmented in a dose dependent manner and reached a plateau level at 0.1 KE/ml of OK-432 . Furthermore, the alveolar macrophages produced various cytokines, i.e., IL-1, TNF-alpha, and IL-6 after 72 h cultivation with 0.1 KE/ml of OK-432 . Based on the in vitro experiments, six patients with intrapulmonary metastatic BAC were therefore treated by the inhalation of aerosolized OK-432 . All 6 patients inhaled aerosolized OK-432 (0.1 KE/ml) twice a day for 5 days . The inhalation therapy regimen was repeated either weekly or monthly unless the tumor markedly progressed . No adverse events were observed in any patients . Either an augmentation of antitumor cytotoxicity or TNF-alpha production by the alveolar macrophages was observed in the two of three patients examined . CONCLUSION: OK-432 inhalation therapy was found to be safe and thus warrants further investigation to determine its clinical effectiveness for BAC. Antonie Van Leeuwenhoek, 2000 Jan, 77(1), 21 - 30 A role for Lewis a antigens on salivary agglutinin in binding to Streptococcus mutans; Ligtenberg AJ et al.; Streptococcus mutans is a major etiological agent in dental caries . Salivary agglutinin is one of the main salivary components binding to S . mutans . To learn more about the interaction of salivary agglutinin with S . mutans, parotid, submandibular, sublingual and palatal saliva samples were incubated with S . mutans suspension . Both depleted saliva samples and bacterial extracts were analyzed by SDS-PAGE and immunoblotting . Salivary agglutinin was present in all types of glandular saliva and in all cases bound to S . mutans, also to PC337C, a P1 mutant of S . mutans . Agglutinin was separated by SDS-PAGE under reducing and non-reducing conditions and then transferred to nitrocellulose . Non-reduced agglutinin bound S . mutans, but reduced agglutinin did not . Adhesion of S . mutans to agglutinin-coated microplates was inhibited by amine-containing components, 1 M NaCl or KCl and EDTA . Adhesion decreased with decreasing pH with no adhesion below pH 5.0 . These data suggest that calcium-dependent electrostatic interactions play a role in binding . By immunoblotting was demonstrated that blood group antigens and Lewis antigens were present on agglutinin . Synthetic blood group antigens and Lewis antigens covalently coupled to polyacrylamide were tested for binding to S . mutans . Only Le(a)(Gal beta 1,3(Fuc alpha 1,4)GlcNAc) bound to S . mutans, whereas the blood group antigens Le(b), Le(x), Le(y), H1, H2, A, B and sialylated Le(a) did not . Lea without galactose (Fuc alpha 1,4GlcNAc) still bound to S . mutans, but Le(a) without fucose (Gal beta 1,3GlcNAc) did not . Binding of agglutinin to S . mutans was not inhibited by Le(a) . In conclusion, S . mutans can bind to Le(a) carbohydrate epitopes in which the fucose is an essential residue . Le(a) carbohydrate epitopes are present on salivary agglutinin but play no major role in binding. Can J Microbiol, 1999 Dec, 45(12), 1027 - 32 Effects of iron and manganese availability on growth and production of superoxide dismutase by Streptococcus suis; Niven DF et al.; A complex medium supported good growth of the type strain of Streptococcus suis irrespective of the presence or absence of a high concentration (1 microM) of the iron chelating agent, ethylenediamine di-o-hydroxyphenylacetic acid . Good growth was also obtained using a complex medium that had been treated with Chelex-100 to reduce the iron content, but only if this medium was supplemented with manganese; supplementation with iron had little effect . Collectively, these results indicate that S . suis requires manganese, but not iron, for growth . Polyacrylamide gel electrophoresis of cell extracts followed by activity staining revealed the presence of a single manganese-cofactored superoxide dismutase; activity staining and enzyme assays revealed that manganese availability during growth affected the activity of the superoxide dismutase in cell extracts . The results are discussed with respect to the capacity of S . suis to avoid damage by reactive oxygen species. J Clin Microbiol, 2000 Mar, 38(3), 1156 - 60 Efficacy of antimicrobial treatments and vaccination regimens for control of porcine reproductive and respiratory syndrome virus and Streptococcus suis coinfection of nursery pigs; Halbur P et al.; Seventy-six, crossbred, porcine reproductive and respiratory syndrome virus (PRRSV)-free pigs were weaned at 12 days of age and randomly assigned to seven groups of 10 to 11 pigs each . Pigs in group 1 served as unchallenged controls . Pigs in groups 2 to 7 were challenged intranasally with 2 ml of high-virulence PRRSV isolate VR-2385 (10(4.47) 50% tissue culture infective doses per 2 ml) on day 0 of the study (30 days of age) . Seven days after PRRSV challenge, pigs in groups 2 to 7 were challenged intranasally with 2 ml of Streptococcus suis serotype 2 (10(8.30) CFU/2 ml) . Group 2 pigs served as untreated positive controls . Antimicrobial treatments included daily intramuscular injection with 66,000 IU of procaine penicillin G per kg of body weight on days 8 to 10 (group 3), drinking water medication with 23.1 mg of tiamulin per kg during days 8 to 10 (group 4), and daily intramuscular injection of 5.0 mg of ceftiofur hydrochloride per kg on days 8 to 10 (group 5) . Vaccination regimens included two intramuscular doses of an autogenous killed S . suis vaccine (group 6) prior to S . suis challenge or a single 2-ml intramuscular dose of an attenuated live PRRSV vaccine (group 7) 2 weeks prior to PRRSV challenge . Mortality was 0, 63, 45, 54, 9, 40, and 81% in groups 1 to 7, respectively . Ceftiofur treatment was the only regimen that significantly (P < 0 . 05) reduced mortality associated with PRRSV and S . suis coinfection . The other treatments and vaccinations were less effective . We conclude that ceftiofur administered by injection for three consecutive days following S . suis challenge was the most effective regimen for minimizing disease associated with PRRSV and S . suis coinfection. J Clin Microbiol, 2000 Mar, 38(3), 977 - 86 Identification of the major Spanish clones of penicillin-resistant pneumococci via the Internet using multilocus sequence typing; Zhou J et al.; Multilocus sequence typing was used to characterize isolates of the major Spanish clones of penicillin-resistant and multiple-antibiotic-resistant Streptococcus pneumoniae . Isolates of the multidrug-resistant Spanish serotype 23F clone and serotype variants of this clone either had identical allelic profiles or their allelic profiles differed from this typical allelic profile at only one of the seven housekeeping loci . Similarly, isolates of the Spanish serotype 6B and 14 clones and the penicillin-resistant serotype 9V clone (and serotype variants of this clone) each had the same allelic profiles or profiles that differed at a single locus . Multilocus sequence typing therefore allows resistant pneumococci to be assigned to the Spanish clones if they have the typical allelic profile of the clone or if their profiles differ from that profile at a single locus . A few resistant isolates that had allelic profiles typical of that of a Spanish clone or whose profiles differed from that of the typical profile at only a single locus possessed penicillin-binding protein pbp1a, pbp2b, or pbp2x genes that differed from those that are characteristic of the clone . In most cases these isolates could be assigned as variant members of the clone . Since almost all serotype 9V isolates have very similar genotypes, independently emerging penicillin-resistant clones of this serotype will inevitably appear to be similar by molecular typing procedures . Analysis of the pbp genes, in addition to multilocus sequence typing (or any other molecular typing procedure), is therefore required to assign isolates unambiguously to the penicillin-resistant Spanish serotype 9V clone. J Oral Pathol Med, 2000 Jan, 29(1), 26 - 32 Lack of association between Streptococcus oralis and recurrent aphthous stomatitis; Riggio MP et al.; In the present study, the potential involvement of Streptococcus oralis in the aetiology of recurrent aphthous stomatitis (RAS) was investigated using the polymerase chain reaction (PCR) . Biopsies from 28 RAS patients were analysed, in addition to 20 oral lichen planus (OLP) and 13 normal biopsies that were used as controls . PCR was carried out using a primer pair that targets the D-alanine:D-alanine ligase gene and detects DNA from both S . oralis and the closely related species Streptococcus mitis . Discrimination between these two species was achieved by digestion of PCR products with the restriction endonucleases HaeIII and HindIII, which both give distinct restriction profiles for each species . S . oralis DNA was detected in 8 of 28 (29%) RAS samples, 10 of 20 (50%) OLP samples and 6 of 13 (46%) normal samples . These results suggest that S . oralis is not of primary aetiological significance in RAS. J Heart Valve Dis, 2000 Jan, 9(1), 131 - 4 Preliminary experience with Silzone-coated St . Jude medical valves in acute infective endocarditis; Bertrand S et al.; BACKGROUND AND AIM OF THE STUDY: The rate of recurrent postoperative endocarditis after valve replacement in early-stage acute infective endocarditis is extremely high . Metallic silver coating of the sewing ring may improve the short- and long-term outcome after valve implantation . This report details our experience with the St . Jude Medical Silzone prosthesis in early surgical treatment of acute infective endocarditis . METHODS: Ten patients (mean age 66.4 years) referred for native valve or prosthetic valve endocarditis were operated on between April 1998 and June 1999 . The microorganisms responsible for the acute infection were Staphylococcus (n = 1), Streptococcus (n = 1) and Pseudomonas aeruginosa (n = 1); blood cultures remained negative in two cases . The indication for surgical treatment was related to hemodynamic condition (n = 5), a major cerebral event (stroke; n = 1), annulus abscess (n = 1), and echocardiographic evidence of large cuspal vegetations (n = 3) . All patients had received preoperative intravenous antibiotics (mean 7.8 days) . Four mitral, five aortic valve replacements, and one double mitral-aortic valve replacement, were performed after extensive debridement of the infected and necrotic tissues . Mean duration of postoperative antibiotic treatment was 32.3 days . Postoperative follow up (mean 6 months; range: 2-14.2 months) was 100% complete, and included prospective repeated transthoracic echocardiography at one week, and one, six and 12 months postoperatively . RESULTS: One patient died early in the immediate postoperative period from pneumonia and major hypoxemia . All other patients are symptom-free, without evidence of recurrent infection and perivalvular leak . CONCLUSION: Although these early results with the St . Jude Medical Silzone prosthesis require confirmation by more extensive studies, they infer that silver coating of the sewing ring may dramatically improve management of patients with active endocarditis. Kansenshogaku Zasshi, 2000 Jan, 74(1), 1 - 5 Characterization of Streptococcus pneumoniae strains isolated from systemic infections in children; Ohkusu K et al.; Twenty-three cases of systemic pneumococcal infection diagnosed from October 1988 to September 1998 were analyzed retrospectively in order to characterize the epidemiology of systemic pneumococcal infections . The clinical diagnosis of those cases were 8 pneumonia, 8 meningitis, 3 septicemia, 3 septic arthritis, and 1 peritonitis . The patients ranged in age from 6 months to 21 years old (mean +/- SD = 3 years, 6 months +/- 5 years, 2 months), and 61% of the patients were younger than 24 months . Resistance to penicillin G (PCG) was detected in 57% of all cases . Resistance to cefotaxime (CTX), imipenem (IPM), erythromycin (EM), and clindamycin (CLDM) was 33%, 9%, 70%, and 65%, respectively . Of the 13 isolates resistant to PCG, 2 were resistant to IPM, 11 to EM and 11 to CLDM . Serotyping was performed on 17 isolates . The identified serotypes were 19 (6 isolates), 6 (5 isolates), 23 (3 isolates), 14 (2 isolates), and 5 (1 isolate) . Eleven isolates resistant to PCG were limited to serotypes 6, 19, or 23 . One patient had a recurrent episode of bacteremic pneumonia 7 months after the first episode . Streptococcus pneumoniae isolates from both episodes were compared by serotyping and pulsed-field gel electrophoresis with restriction digestion, and were confirmed as the same strain. Br Med Bull, 1999, 55(1), 259 - 76 Antibacterial resistance in the intensive care unit: mechanisms and management; Elliott TS et al.; The incidence of multiple antimicrobial resistance of bacteria which cause infections in the intensive care unit is increasing . These include methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci, penicillin-resistant Streptococcus pneumoniae and cephalosporin and quinolone resistant coliforms . More recently, pan antibiotic resistant coliforms, including carbapenems, have emerged . The rapidity of emergence of these multiple antibiotic-resistant organisms is not being reflected by the same rate of development of new antimicrobial agents . It is, therefore, conceivable that patients with serious infections will soon no longer be treatable with currently available antimicrobials . Strict management of antibiotic policies and surveillance programmes for multiple resistant organisms, together with infection control procedures, need to be implemented and continuously audited . As intensive care units provide a nidus of infection for other areas within hospitals, this is critically important for prevention of further spread and selection of these resistant bacteria. Epidemiol Infect, 1999 Dec, 123(3), 383 - 8 Nasopharyngeal colonization of infants in southern India with Streptococcus pneumoniae; Jebaraj R et al.; To investigate the dynamics of nasopharyngeal colonization with Streptococcus pneumoniae, and to determine the prevalent serogroups/types (SGT) and their antimicrobial susceptibility, we studied 100 infants attending our well-baby clinic . Nasopharyngeal swab specimens were obtained at 6, 10, 14, 18 and 22 weeks and at 9 and 18 months of age and submitted for culture, serotyping and antimicrobial susceptibility testing of S . pneumoniae . Colonization with pneumococcus was seen on at least one occasion in 81 infants . The median age of acquisition was 11 weeks and the median duration of carriage was 1 3 months . The common SGTs identified were 6, 19, 14 and 15 . SGT 1, which was a common invasive isolate in children in our hospital during this period, was not isolated from these children . Sequential colonization by 2, 3 or 4 SGTs was observed in 18, 5 and 2 children, respectively . Resistance to penicillin, chloramphenicol, cotrimoxazole and erythromycin was observed in 0, 13 (6%) 11 (5 %) and 5 (3 %) isolates, respectively . There was a significant difference in susceptibility to cotrimoxazole between colonizing and invasive isolates (5 % vs . 40 %, P<0.0001). Pediatr Infect Dis J, 2000 Feb, 19(2), 134 - 8 Human antibodies to pneumococcal surface protein A in health and disease; Virolainen A et al.; BACKGROUND: Diseases caused by Streptococcus pneumoniae have a high impact in young children whose ability to mount antibodies to capsular polysaccharides is impaired . Pneumococcal surface protein A (PspA) is a potential vaccine candidate for this age group . METHODS: We used Western blot analysis and enzyme immunoassay to study human sera of healthy adults from Alabama (n = 20) and from Finland (n = 21), healthy children from Finland (n = 20) and ill children from Finland, those with pneumococcal invasive infection (n = 26) and those with nonpneumococcal invasive infection (n = 26) . RESULTS: Human antibodies to PspA exhibited strong cross-reactivity among different pneumococcal strains . The geometric mean titer of IgG antibody to PspA in sera from 21 healthy adults was 4,040, from ten 3-year-old healthy children 1,080 and from ten 2-month-old healthy children 1,650 . The geometric mean titer of PspA antibody of acute phase sera of children with invasive pneumococcal disease was 140, significantly (P < 0.001) lower than the respective value, 1,020, for children with infection caused by other bacteria . CONCLUSIONS: We demonstrate for the first time the existence of antibodies to PspA in human sera in health and disease . The findings in ill children suggest that antibodies to PspA might play a role in protection against pneumococcal disease. Clin Transplant, 2000 Feb, 14(1), 61 - 5 Immunization of renal transplant recipients with pneumococcal polysaccharide vaccine; Kazancioglu R et al.; BACKGROUND: Streptococcus pneumoniae, a common pathogen leading to pneumonia, is a cause of morbidity and mortality in immunosuppressed patients . Vaccination against this agent can be recommended for immunosuppressed patients, including those with chronic renal failure, nephrotic syndrome and renal transplant recipients; however, a diminished immune response and loss of protective antibodies have been observed . PATIENTS AND METHODS: In our prospective study, the efficacy and side effects of polyvalent pneumococcal vaccination were investigated in renal transplant recipients . A total of 21 patients (6 female, 15 male) with well-functioning renal allografts, who had transplant surgery at least 2 months before, were included in the study . The patients were stratified according to the immunosuppressive protocol and 8 received double, while 13 received triple, immunosuppressive agents . After obtaining basal serum samples, all cases were vaccinated with the 0.5 mL intramuscular administration of polyvalent polysaccharide pneumococcal vaccine (Pneumo 23 Pasteur Merieux, lot No: K 1131) . RESULTS: Following a mean of 6 wk in all patients and also a mean of 12 wk in 12 patients, serum samples were again obtained to measure pneumococcal antibodies . Antibody titers following 6 and 12 wk of vaccination were significantly higher, as compared with basal values in all patients, except one . These titers did not show any statistically significant difference between double and triple therapies . There was no significant difference between the 12th and 6th wk postvaccination antibody titers . No systemic or local adverse effects were observed . CONCLUSION: Pneumococcal vaccination is safe and effective in patients with well-functioning renal allografts, at least in the short term . This vaccination policy may be useful for preventing invasive pneumococcal disease in immunosuppressed patients. J Oral Sci, 1999 Dec, 41(4), 169 - 72 The phenomenon of salivary protein adsorption onto Streptococcus mitis ATCC 903 cells; Tamura M et al.; It is thought that salivary proteins on bacterial cells have some influence on the adhesion of oral bacteria onto the surface of oral tissues and on bacterial aggregation . Initially, this study sought to examine the phenomenon of salivary protein adsorption to the surface of Streptococcus mitis ATCC 903 using 3H-labeled salivary proteins . We investigated the effects of hydrophobic level and lectin-ligand binding on adsorption of salivary proteins, and also the influence of cell surface components on adsorption . The results showed that the adsorption of salivary proteins was clearly reduced by the presence of Tween 20, LiCl, NaSCN, Hexadecane and some sugars . The adsorption was also affected by the pH level, and protease treatment or heat treatment of cells also decreased the volume of adsorbed proteins . Although the adsorption of proteins onto heat-treated cells was dramatically reduced by the presence of LiCl and NaSCN, the presence of sugars had little influence on this adsorption . These findings suggest that the main adsorption of salivary proteins is due to hydrophobic factors, and the heat-sensitive surface components of the cells had some relation to lectin-ligand binding . Therefore, it appears important to study the adsorption of salivary proteins onto cells since the salivary proteins on bacterial cells play an important role in their adherence to the saliva-coated oral tissues and bacterial aggregation. J Bacteriol, 2000 Mar, 182(6), 1529 - 40 The RofA binding site in Streptococcus pyogenes is utilized in multiple transcriptional pathways; Granok AB et al.; Understanding the regulation of adhesins defines a pathogenic bacterium's interaction with the local environment within the host . In certain strains of Streptococcus pyogenes, transcription of prtF, the gene which encodes the fibronectin-binding adhesin protein F, is activated by RofA under anaerobic conditions . RofA binds specifically to DNA in its target promoters and autoregulates its own expression . In this study, we have used DNase I protection assays to further investigate the interaction of RofA with its target promoters . In the region between rofA and the gene which encodes protein F (prtF), RofA binds to two distinct sites: a smaller site (17 bp) adjacent to the rofA promoter, and a larger site (40 bp) adjacent to the prtF promoter . Analysis of fusions to a novel reporter gene whose product consists of the fusion of the N-terminal secretion domain of protein F with the C-terminal enzymatic domain of the enterococcal alkaline phosphatase (PhoZ) revealed that the small RofA binding site had no direct role in control of prtF transcription but contributed to regulation of rofA . Comparison in several strains representing different patterns of prtF expression indicated that the larger site was required for activation of rofA and of prtF in all strains by both RofA-dependent and -independent pathways . Thus, it would appear that a common recognition sequence provides separate entries to a final common pathway in S . pyogenes virulence gene expression . The identification of multiple RofA-like proteins and promoters with RofA binding sites implies the existence of a widespread interacting regulatory network. Drugs Aging, 1999, 15 Suppl 1, 11 - 9 Epidemiology of pneumococcal infections in the elderly; Butler JC et al.; The risk of invasive Streptococcus pneumoniae infection (primarily bacteraemia and meningitis) is greatest among the very young and the very old . Persons in certain racial groups, including African-Americans, American Indians, Native Alaskans and Australian Aborigines, are also at increased risk of disease . Other factors that appear to increase the risk of pneumococcal infection are lower socioeconomic status, recent infection with influenza and possibly other viral respiratory tract infections, chronic medical conditions, and immunosuppressive medications . Reported annual incidences of invasive pneumococcal disease among persons aged > or = 65 years in North America and Europe range from 25 to 90 cases/100,000 persons . In the US and Canada, these rates represent between 15,000 and 30,000 cases annually among the elderly . Mortality caused by pneumococcal infections is highest among the elderly, with nearly 1 in 5 cases resulting in death . Worldwide, S . pneumoniae is the leading cause of community-acquired pneumonia requiring hospitalisation . The high fatality rates, as well as recent outbreaks of pneumococcal infection among unvaccinated nursing home residents and the emergence of drug-resistant pneumococcal strains, highlight the importance of preventing invasive infection by vaccination. J Vet Diagn Invest, 2000 Jan, 12(1), 21 - 7 Coinfection by porcine circoviruses and porcine parvovirus in pigs with naturally acquired postweaning multisystemic wasting syndrome; Ellis JA et al.; Postweaning multisystemic wasting syndrome (PMWS) is an emerging disease in swine . Recently, the disease has been reproduced with inocula containing a newly described porcine circovirus (PCV), designated PCV 2, and porcine parvovirus (PPV) . In order to determine if these viruses interact in naturally acquired PMWS, affected tissues from field cases were examined by immunohistochemistry (IHC) and polymerase chain reaction (PCR) for PCV 2 and PPV, as well as by PCR for the other recognized porcine circovirus, PCV 1 . Porcine circovirus 2 was detected by PCR or IHC in affected fixed or frozen tissues from 69 of 69 cases of PMWS collected over 3 years from 25 farms . Porcine parvovirus was detected in 12 of the same cases, and PCV 1 was detected in 9 of 69; however, an apparent decrease was found in the sensitivity of the PCRs used to detect the latter 2 viruses when fixed tissue from the same cases were compared with the use of frozen tissues . Porcine circovirus 2 was not detected by PCR in affected tissues from 16 age-matched pigs that had Streptococcus suis-associated disease . Electron microscopic examination of plasma pooled from 15 pigs with PMWS revealed the presence of PCV and PPV, whereas these viruses were not observed in pooled plasma from 5 age-matched clinically normal pigs . These results confirm and extend previous findings documenting a consistent association of PCV 2 with PMWS . As well, infection by PPV or PCV 1 or both may be an important cofactor in the pathogenesis of some, but apparently not all, cases of PMWS. Mol Med Today, 2000 Mar, 6(3), 125 - 32 Superantigens - powerful modifiers of the immune system; Fraser J et al.; Superantigens are powerful microbial toxins that activate the immune system by binding to class II major histocompatibility complex and T-cell receptor molecules . They cause a number of diseases characterized by fever and shock and are important virulence factors for two human commensal organisms, Staphylococcus aureus and Streptococcus pyogenes, as well as for some viruses . Their mode of action and variation around the common theme of over-stimulating T cells, provides a rich insight into the constant battle between microbes and the immune system. FEMS Microbiol Lett, 2000 Mar 1, 184(1), 17 - 21 Signal sequence and alanine-rich region of streptococcal protein antigen A of Streptococcus sobrinus can direct localization of alkaline phosphatase to the periplasm of Escherichia coli; Holt RG et al.; Streptococcal protein antigen A (SpaA) of Streptococcus sobrinus is expressed on the surface of cells and extracellularly . TnphoA which lacks signals for transcription and membrane transport of Escherichia coli alkaline phosphatase was used to analyze the sequences necessary for transport of a SpaA/PhoA fusion protein across the cytoplasmic membrane to the periplasm of E . coli cells . Of 15 alkaline phosphatase-producing isolates analyzed, all were found to localize more than 85% of the SpaA/PhoA hybrid protein to the periplasm of E . coli cells . From DNA sequence analysis, all were found to have TnphoA inserted into an identical site . The insertion site of TnphoA was downstream from the coding sequence that generates four tandemly repeated alanine-rich sequences of 82 amino acid residues . These results suggest that in addition to the signal sequence, mature protein sequences containing alanine-rich repeat sequences may play a role in the export of the SpaA protein across a bacterial membrane. Infect Immun, 2000 Mar, 68(3), 1569 - 73 Immune responses to specific antigens of Streptococcus pneumoniae and Moraxella catarrhalis in the respiratory tract; Samukawa T et al.; Streptococcus pneumoniae and Moraxella catarrhalis are two common respiratory pathogens, colonizing as many as 54 and 72% of children, respectively, by 1 year of age . The immune responses to surface protein A of S . pneumoniae (PspA) and the high-molecular-weight outer membrane protein of M . catarrhalis (UspA) in the sera of various age groups in the general population and in the nasopharynges of 30 children monitored from birth through 1 year of age were evaluated . Immunoglobulin G (IgG) was the dominant serum antibody to PspA and UspA . Whereas the serum antibody response to PspA peaked in childhood, the antibody response to UspA peaked in adulthood . In the first 2 years of life, comparable amounts of IgM and IgG antibodies to both proteins were observed . In older persons, IgG antibodies to both antigens predominated over IgM antibodies . The levels of IgA antibody to these antigens in serum remained low during the first 2 years of life . The levels of IgM antibody to the two antigens in serum exceeded the levels of IgA antibody to the same two antigens throughout life . Although IgA was the dominant antibody to PspA and UspA in airway secretions, it was detected in a minority of the children (3 of 15 for PspA and 0 of 15 for UspA) . Even the majority of the children previously colonized with these pathogens lacked antibody to them in their secretions. Infect Immun, 2000 Mar, 68(3), 1557 - 62 Relative roles of pneumolysin and hydrogen peroxide from Streptococcus pneumoniae in inhibition of ependymal ciliary beat frequency; Hirst RA et al.; Ciliated ependymal cells line the ventricular system of the brain and the cerebral aqueducts . This study characterizes the relative roles of pneumolysin and hydrogen peroxide (H(2)O(2)) in pneumococcal meningitis, using the in vitro ependymal ciliary beat frequency (CBF) as an indicator of toxicity . We have developed an ex vivo model to examine the ependymal surface of the brain slices cut from the fourth ventricle . The ependymal cells had cilia beating at a frequency of between 38 and 44Hz . D39 (wild-type) and PLN-A (pneumolysin-negative) pneumococci at 10(8) CFU/ml both caused ciliary slowing . Catalase protected against PLN-A-induced ciliary slowing but afforded little protection from D39 . Lysed PLN-A did not reduce CBF, whereas lysed D39 caused rapid ciliary stasis . There was no effect of catalase, penicillin, or catalase plus penicillin on the CBF . H(2)O(2) at a concentration as low as 100 microM caused ciliary stasis, and this effect was abolished by coincubation with catalase . An additive inhibition of CBF was demonstrated using a combination of both toxins . A significant inhibition of CBF at between 30 and 120 min was demonstrated with both toxins compared with either H(2)O(2) (10 microM) or pneumolysin (1 HU/ml) alone . D39 released equivalent levels of H(2)O(2) to those released by PLN-A, and these concentrations were sufficient to cause ciliary stasis . The brain slices did not produce H(2)O(2), and in the presence of 10(8) CFU of D39 or PLN-A per ml there was no detectable bacterially induced increase of H(2)O(2) release from the brain slice . Coincubation with catalase converted the H(2)O(2) produced by the pneumococci to H(2)O . Penicillin-induced lysis of bacteria dramatically reduced H(2)O(2) production . The hemolytic activity released from D39 was sufficient to cause rapid ciliary stasis, and there was no detectable release of hemolytic activity from the pneumolysin-negative PLN-A . These data demonstrate that D39 bacteria released pneumolysin, which caused rapid ciliary stasis . D39 also released H(2)O(2), which contributed to the toxicity, but this was masked by the more severe effects of pneumolysin . H(2)O(2) released from intact PLN-A was sufficient to cause rapid ciliary stasis, and catalase protected against H(2)O(2)-induced cell toxicity, indicating a role for H(2)O(2) in the response . There is also a slight additive effect of pneumolysin and H(2)O(2) on ependymal toxicity; however, the precise mechanism of action and the role of these toxins in pathogenesis remain unclear. Infect Immun, 2000 Mar, 68(3), 1450 - 6 CpG oligodeoxynucleotides act as adjuvants for pneumococcal polysaccharide-protein conjugate vaccines and enhance antipolysaccharide immunoglobulin G2a (IgG2a) and IgG3 antibodies; Chu RS et al.; Pneumococcal polysaccharide-protein conjugate vaccines elicit antipolysaccharide antibodies, but multiple doses are required to achieve protective antibody levels in children . In addition, the immunogenicity of experimental multivalent pneumococcal conjugate vaccines varies with different polysaccharide serotypes . One strategy to improve these vaccines is to incorporate an adjuvant to enhance their immunogenicity . Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are adjuvants that promote T-cell and T-dependent antibody responses to protein antigens, but it has been unclear whether CpG ODN can enhance polysaccharide-specific antibody responses . The present studies demonstrate significant adjuvant activity of CpG ODN for antibody responses against Streptococcus pneumoniae polysaccharide types 19F and 6B induced by conjugates of 19F and 6B with the protein carrier CRM(197) . BALB/c ByJ mice were injected with 19F-CRM(197) or 6B-CRM(197) with or without CpG ODN, and sera were tested for anti-19F or anti-6B antibodies by enzyme-linked immunosorbent assay . The polysaccharide-specific antibody response to 19F-CRM(197) alone was predominantly of the immunoglobulin G1 (IgG1) and IgM isotypes, but addition of CpG ODN markedly increased geometric mean titers of total anti-19F antibody (23-fold), anti-19F IgG2a (26-fold), and anti-19F IgG3 (>246-fold) . The polysaccharide-specific antibody response to 6B-CRM(197) alone consisted only of IgM, but addition of CpG ODN induced high titers of anti-6B IgG1 (>78-fold increase), anti-6B IgG2a (>54-fold increase), and anti-6B IgG3 (>3,162-fold increase) . CpG ODN also increased anti-CRM(197) IgG2a and IgG3 . Adjuvant effects were not observed with control non-CpG ODN . Thus, CpG ODN significantly enhance antipolysaccharide IgG responses (especially IgG2a and IgG3) induced by these glycoconjugate vaccines. Infect Immun, 2000 Mar, 68(3), 1374 - 82 Genetic relationships between clinical isolates of Streptococcus pneumoniae, Streptococcus oralis, and Streptococcus mitis: characterization of "Atypical" pneumococci and organisms allied to S . mitis harboring S . pneumoniae virulence factor-encoding genes; Whatmore AM et al.; The oral streptococcal group (mitis phylogenetic group) currently consists of nine recognized species, although the group has been traditionally difficult to classify, with frequent changes in nomenclature over the years . The pneumococcus (Streptococcus pneumoniae), an important human pathogen, is traditionally distinguished from the most closely related oral streptococcal species Streptococcus mitis and Streptococcus oralis on the basis of three differentiating characteristics: optochin susceptibility, bile solubility, and agglutination with antipneumococcal polysaccharide capsule antibodies . However, there are many reports in the literature of pneumococci lacking one or more of these defining characteristics . Sometimes called "atypical" pneumococci, these isolates can be the source of considerable confusion in the clinical laboratory . Little is known to date about the genetic relationships of such organisms with classical S . pneumoniae isolates . Here we describe these relationships based on sequence analysis of housekeeping genes in comparison with previously characterized isolates of S . pneumoniae, S . mitis, and S . oralis . While most pneumococci were found to represent a closely related group these studies identified a subgroup of atypical pneumococcal isolates (bile insoluble and/or "acapsular") distinct from, though most closely related to, the "typical" pneumococcal isolates . However, a large proportion of isolates, found to be atypical on the basis of capsule reaction alone, did group with typical pneumococci, suggesting that they have either lost capsule production or represent as-yet-unrecognized capsular types . In contrast to typical S . pneumoniae, isolates phenotypically identified as S . mitis and S . oralis, which included isolates previously characterized in taxonomic studies, were genetically diverse . While most of the S . oralis isolates did fall into a well-separated group, S . mitis isolates did not cluster into a well-separated group . During the course of these studies we also identified a number of potentially important pathogenic isolates, which were frequently associated with respiratory disease, that phenotypically and genetically are most closely related to S . mitis but which harbor genes encoding the virulence determinants pneumolysin and autolysin classically associated with S . pneumoniae. Infect Immun, 2000 Mar, 68(3), 1215 - 21 Role of group A streptococcal virulence factors in adherence to keratinocytes; Darmstadt GL et al.; To evaluate the role of putative group A streptococcal virulence factors in the initiation of skin infections, we compared the adherence of a wild-type M49-protein skin-associated strain to that of a series of 16 isogenic mutants created by insertional inactivation of virulence genes . None of the mutants, including the M-protein-deficient (emm mutant) strain, displayed reduced adherence to early-passage cultured human keratinocytes, but adherence of the mutant lacking hyaluronic acid capsule expression (has mutant) was increased 13-fold . In contrast, elimination of capsule expression in M2-, M3-, and M18-protein has mutants increased adherence only slightly (1.3- to 2.3-fold) compared to their respective wild-type strains . A mutant with inactivation of both emm and has displayed high-level adherence (34.9 +/- 4.1%) equal to that of the has mutant strain (40.7 + 8.0%), confirming the lack of involvement of M49 protein in attachment . Moreover, adherence of the M49-protein-deficient (emm mutant) and wild-type strains was increased to the same level (57 and 55%, respectively) following enzymatic digestion of their hyaluronic acid capsule . Adherence of mutants lacking oligopeptide permease (Opp) expression was increased 3.8- to 5.5-fold, in association with decreased cell-associated hyaluronic acid capsule . Finally, soluble CD46 failed to inhibit adherence of M49- and M52-serotype skin strains . We conclude that (i) bacterial M protein and keratinocyte CD46 do not mediate adherence of M49 skin-associated Streptococcus pyogenes to epidermal keratinocytes, (ii) hyaluronic acid capsule impedes the interaction of bacterial adhesins with keratinocyte receptors, (iii) modulation of capsule expression may be important in the pathogenesis of skin infections, and (iv) the molecular interactions in attachment of skin strains of S . pyogenes to keratinocytes are unique and remain unidentified. Infect Immun, 2000 Mar, 68(3), 1019 - 25 Allele substitution of the streptokinase gene reduces the nephritogenic capacity of group A streptococcal strain NZ131; Nordstrand A et al.; To investigate the role of allelic variants of streptokinase in the pathogenesis of acute poststreptococcal glomerulonephritis (APSGN), site-specific integration plasmids were constructed, which contained either the non-nephritis-associated streptokinase gene (skc5) from the group C streptococcal strain Streptococcus equisimilis H46A or the nephritis-associated streptokinase gene (ska1) from the group A streptococcal nephritogenic strain NZ131 . The plasmids were introduced by electroporation and homologous recombination into the chromosome of an isogenic derivative of strain NZ131, in which the streptokinase gene had been deleted and which had thereby lost its nephritogenic capacity in a mouse model of APSGN . The introduction of a non-nephritis-associated allelic variant of streptokinase did not rescue the nephritogenic capacity of the strain . The mutant and the wild-type strains produced equivalent amounts of streptokinase . Complementation of the ska deletion derivative with the original ska allele reconstituted the nephritogenicity of wild-type NZ131 . The findings support the hypothesis that the role of streptokinase in the pathogenesis of APSGN is related to the allelic variant of the protein. Clin Ther, 2000 Jan, 22(1), 29 - 39 Antimicrobial therapy of acute otitis media: review of treatment recommendations; Aronovitz GH; OBJECTIVE: This paper reviews 3 previously published articles that provided recommendations for antimicrobial therapy of acute otitis media (AOM) and combines them to provide revised recommendations . BACKGROUND: AOM is one of the most common pediatric infections requiring a prescription for an antimicrobial agent . The optimal approach to treatment of AOM requires early, efficacious, and practical therapy . Several experts and organizations have developed recommendations for the management of AOM, but the number of these may overwhelm the busy primary care practitioner . A MEDLINE search of the pediatric and infectious disease literature on AOM treatment recommendations was used to select 3 representative, previously published articles for this review . When selecting an agent, physicians should consider in vitro activity, particularly against drug-resistant Streptococcus pneumoniae; pharmacokinetics; adverse events; palatability of the suspension; and cost . In addition, physicians' clinical experience is an important determinant . CONCLUSIONS: Amoxicillin is recommended as the first-line agent to treat uncomplicated AOM . For clinical treatment failures after 3 days of amoxicillin, recommended antimicrobial agents include oral amoxicillin/clavulanate, cefuroxime axetil, cefprozil, cefpodoxime proxetil, and intramuscular (i.m.) ceftriaxone . I.m . ceftriaxone should be reserved for severe cases or patients in whom noncompliance is expected . Tympanocentesis for identification of pathogens and susceptibility to antimicrobial agents is recommended for selection of third-line agents. Clin Ther, 2000 Jan, 22(1), 2 - 14 Comparison of bacteriologic eradication of Streptococcus pneumoniae by clarithromycin and reports of increased antimicrobial resistance; Gotfried MH; OBJECTIVE: To determine whether reported increases in Streptococcus pneumoniae resistance, as determined by in vitro antimicrobial susceptibility testing, correlate with the clinical efficacy of clarithromycin in treating patients with acute exacerbations of chronic bronchitis (AECB) or community-acquired pneumonia (CAP) . BACKGROUND: Surveillance data on antimicrobial resistance suggest that the overall rate of S . pneumoniae resistance in vitro in the United States has increased to approximately 45% during the past decade . S . pneumoniae is showing increased resistance to penicillin, other beta-lactams, and macrolides . Despite this increased resistance, the clinical efficacy of clarithromycin does not appear to be diminished to the degree suggested by reported resistance rates . The author examined several studies of clarithromycin in patients with AECB or CAP that demonstrate S . pneumoniae eradication rates in vivo of approximately 92% . The discordance between reported increases in resistance of S . pneumoniae isolates in vitro and the eradication rate with clarithromycin in vivo is discussed in light of 5 observations . RESULTS: First, surveillance data on S . pneumoniae resistance rates to clarithromycin may be overestimated . Second, efflux mutant strains may not be clinically resistant . Third, host immune defenses play a role in treatment outcomes . Fourth, in vitro resistance may not correlate with in vivo clinical success . Finally, clarithromycin and its active metabolite, 14-OH-clarithromycin, attain high concentrations in patients . CONCLUSION: Reported increases in the prevalence of S . pneumoniae resistance do not appear to have had proportional effects on the clinical efficacy of clarithromycin in the treatment of patients with AECB or CAP caused by S . pneumoniae. Ostomy Wound Manage, 1999 Oct, 45(10), 50 - 4, 56-8 The effects of UVC irradiation on group A streptococcus in vitro; Sullivan PK et al.; Streptococcus pyogenes (group A streptococcus--GAS) is a common cause of necrotizing fasciitis (NF)--a severe infection of the subcutaneous soft tissue . The purpose of this study was to determine if the topical therapy ultraviolet light C (UVC) is effective in killing GAS in vitro and to evaluate the most effective treatment parameters for use with UVC therapy . Five replications of GAS at 10(8) organisms/mL were plated . The cultures were treated with a UVC light 1 inch from the surface . Irradiation times were as follows: 0, 2, 3, 4, 5, 15, 30, 45, 60, 90, 120, and 180 seconds . Bacterial cultures were incubated and colony counts performed . A second set of GAS cultures were exposed to UVC for 30, 90, and 120 seconds either once daily (qd) or twice daily (bid) . Kill rates were 99.9% for GAS at 4 seconds to 180 seconds . Kill rates of 99.9% were also obtained at 30 seconds and 90 seconds when UVC treatment was given either qd or bid . This data indicates that UVC is bactericidal for GAS at times as short as 4 seconds . In addition, UVC treatment was not effective when administered through thin film dressings. J Endod, 1999 Oct, 25(10), 653 - 9 Bacterial microleakage of Cavit, IRM, TERM, and Fermit: a 21-day in vitro study; Deveaux E et al.; The aim of our study was to evaluate the leakage of four cements (Cavit, IRM, TERM, and Fermit) using a two-compartment model system and Streptococcus sanguis as bacterial marker . Access cavities in premolars were filled with cement and the teeth immersed in culture medium in the model system . Half of the teeth were thermocycled on day 2 . Bacterial percolation into the upper compartment was measured at regular intervals (days 2, 7, 14, and 21) . Cement thickness was measured at the end of the study . In the nonthermocycled group, Cavit was more leakproof than the other cements at day 2 (p = 0.011), than TERM and IRM at day 7 (p = 0.043) . Fermit was more leakproof than IRM at day 7 (p = 0.043) . In the thermocycled group, Cavit was more leakproof than the other cements at day 7 (p = 0.041) . Thermocycling did not significantly affect leakage . Cement thickness averaged 4.1 mm and did not significantly affect leakage . These results should be considered when using cements as temporary fillings. Gen Dent, 1999 Mar-Apr, 47(2), 187 - 90 Antimicrobial activity of cavity disinfectants; Gultz J et al.; The purpose of this study was to determine the antimicrobial activity of four commercially available cavity disinfectants and one prescription mouthwash as they came into contact with bacteria commonly found in the oral cavity . Streptococcus mutans, salivarius, and Actinomyces viscosus were used in the study . Zones of microbial inhibition were measured in millimeters after 48 hours . The results of this study indicate that all of the antimicrobial agents demonstrated activity against the bacteria tested . Consepsis Solution produced the largest zones of inhibition against all three of the bacteria used. Clin Chim Acta, 2000 Feb 25, 292(1-2), 1 - 12 Glutamine synthetase in experimental meningitis: increased ratio of the subunits 3 and 2 may indicate enhanced activity; Smirnov AV et al.; Glutamine synthetase (GS) activity is higher in the neocortex but not in the hippocampal formation of rabbit brain during Streptococcus pneumoniae meningitis compared to the respective brain region of uninfected control animals . One-dimensional polyacrylamide gel electrophoresis (1D-SDS-PAGE) revealed an apparent molecular mass (M(r)) of 44000 Dalton (Da) for GS from rabbit brain . After two-dimensional gel electrophoresis (2D-PAGE), followed by Coomassie-blue staining, GS separated into three distinct spots (S1, S2, S3) . One additional spot (S4) occurred on the immunoblot . All four GS spots exhibited the same M(r) (44000 Da), but differed in their isoelectric points . Densitometric evaluation of the two-dimensional maps revealed a strong increase of optical density (OD) of S3 in the frontal cortex of infected animals . The calculated OD ratio S3/S2 in the frontal cortex from rabbits with meningitis was 1.75+/-0.68 (mean+/-standard deviation) . Compared to controls (0 . 85+/-0.39), this value was significantly increased (p=0.0006) . In the hippocampal formation, the ratio S3/S2 was nearly unchanged during meningitis . It is suggested that the ratio S3/S2 may indicate a neuroprotective feature of rabbit brain during meningitis since neuronal apoptosis occurs only in the dentate gyrus and not in the frontal cortex. Am J Vet Res, 2000 Feb, 61(2), 162 - 6 Comparison of the phenotypes of Streptococcus zooepidemicus isolated from tonsils of healthy horses and specimens obtained from foals and donkeys with pneumonia; Anzai T et al.; OBJECTIVE: To determine whether streptococcal pneumonia is caused by strains of Streptococcus zooepidemicus similar to those obtained from the tonsils of healthy horses . SAMPLE POPULATION: 5 tonsils from healthy horses, 8 tracheal washes and 6 lung specimens from foals with pneumonia, and 5 nasopharyngeal swab specimens from donkeys with acute bronchopneumonia . PROCEDURE: Variable M-like protectively immunogenic SzP proteins of 5 isolates of S . zooepidemicus from each tonsil and clinical specimen were compared, using immunoblots . The SzP gene of 13 isolates representative of various SzP immunoblot phenotypes from 1 healthy horse and 9 horses and donkeys with pneumonia were sequenced and compared . Cell-associated hyaluronic acid concentration and resistance to phagocytosis of some isolates were measured . RESULTS: Tonsils of each healthy horse were colonized by several SzP phenotypes similar to those of foals or donkeys with pneumonia . In contrast, multiple isolates from animals with pneumonia had the same SzP phenotype, indicating infection by a single strain or clone . Analysis of the SzP sequence confirmed that differences in immunoblot phenotype were associated with sequence differences and that several SzP genotypes were in healthy horses and animals with pneumonia . Isolates with high concentrations of cell-associated hyaluronic acid were more resistant to phagocytosis . CONCLUSIONS AND CLINICAL RELEVANCE: An SzP-specific immunoblot is a useful, sensitive measure of diversity among strains of S . zooepidemicus . Single strains with SzP phenotypes similar to those found in tonsils of healthy horses cause pneumonia . Because of the diversity of SzP phenotype and genotype among isolates from animals with pneumonia, SzP phenotype is not an important determinant of invasiveness or epizootic capabilities. J Perinatol, 1999 Jul-Aug, 19(5), 337 - 42 Group B streptococcus: to culture or not to culture? Katz PF, Hibbard JU, Ranganathan D, Meadows W, Ismail M. OBJECTIVE: To determine if universal Group B Streptococcus (GBS) culturing and antibiotic prophylaxis of obstetric patients decreased the incidence of neonatal early-onset GBS sepsis and mortality and maternal chorioamnionitis . STUDY DESIGN: A time series observational study was conducted to compare the cohort of all obstetric patients delivering at the University of Chicago neonatal center from January 1989 through December 1993, before a GBS surveillance policy existed, with the cohort delivering January 1994 through December 1996, after initiation of a GBS policy . Included in the policy were universal GBS cultures at 28 weeks' gestation, antibiotic prophylaxis at the time of labor for all those with positive cultures and for all with risk factors of preterm delivery, preterm premature rupture of membranes, prolonged rupture of membranes greater than 18 hours, and a previous child affected by GBS or maternal fever in labor . Predictor variables were GBS culturing and antibiotic usage; outcome variables were incidence of GBS sepsis and mortality in the neonates and maternal chorioamnionitis . chi-squared and Fisher exact analyses were used with p < 0.05 being significant . RESULTS: Before the GBS policy, there were 16,272 deliveries with a 2.24/1000 deliveries rate of early-onset GBS sepsis (n = 35); after initiating the GBS policy, 9130 deliveries occurred with an early-onset GBS sepsis rate of 2.29/1000 (n = 20) . Early-onset GBS sepsis case fatality rates before and after initiation of the policy were 14.3% and 0%, respectively (p = 0.09) . Antibiotic use almost doubled (relative risk = 1.84; confidence interval, 1.74 to 1.93, p < 0.001) over the two time periods, and the relative risk of chorioamnionitis decreased to 0.95 (confidence interval, 0.73 to 0.99, p = 0.04) . CONCLUSION: Despite universal GBS culturing and very liberal use of antibiotics in labor, we were unable to effect a statistically significant change in the rate of early-onset GBS sepsis or mortality, and there was only a slightly decreased chorioamnionitis rate. Res Vet Sci, 2000 Feb, 68(1), 33 - 9 Determination of intraspecies variations of the V2 region of the 16S rRNA gene of Streptococcus equi subsp . zooepidemicus; Abdulmawjood A et al.; The 16S rRNA gene of 39 S . equi subsp . zooepidemicus strains and two S . equi subsp . equi strains was amplified by polymerase chain reaction and subsequently digested with the restriction enzyme Hinc II . A restriction profile with two fragments with sizes of 1250 bp and 200 bp could be observed for both S . equi subsp . equi strains and for 30 of the 39 S . equi subsp . zooepidemicus strains indicating a sequence variation within the V2 region of the 16S rRNA gene of the remaining nine S . equi subsp . zooepidemicus isolates . A segment of the 16S rRNA gene including the hypervariable V2 region of 11 S . equi subsp . zooepidemicus and two S . equi subsp . equi could be amplified by PCR and sequenced . The sequence of the V2 region of eight S . equi subsp . zooepidemicus strains appeared to be identical or almost identical to the sequence of the two S . equi subsp . equi strains . The sequence of the remaining three S equi subsp . zooepidemicus strains differed significantly from the sequence of S . equi subsp . equi . These differences allowed a division of S . equi subsp . zooepidemicus strains into two 16S rRNA types and might possibly have consequences for the taxonomic position of these phenotypically indistinguishable strains of one subspecies . A molecular typing could additionally be performed by amplification of the gene encoding the 16S-23S rRNA spacer region . A single amplicon of the spacer gene of 1100 bp could be observed for one S . equi subsp . zooepidemicus, an amplicon of 950 bp for two S . equi subsp . equi strains and 10 S . equi subsp . zooepidemicus strains, a amplicon of 780 bp for 27 S . equi subsp . zooepidemicus strains and a single amplicon of 600 bp for one S . equi subsp . zooepidemicus strain . The variations of the V2 region of the 16S rRNA gene and the size variations of the 16S-23S rRNA spacer gene were not related to each other . Both variations could be used for molecular typing of this species, possibly useful in epidemiological aspects . Biopolymers, 2000 Apr 5, 53(4), 281 - 92 Synthesis, microbicidal activity, and solution structure of the dodecapeptide from bovine neutrophils; Raj PA et al.; The dodecapepetide sequence R-L-C-R-I-V-V-I-R-V-C-R with a disulfide bridge between the cysteine residues found in bovine neutrophils was synthesized by solid-phase procedures . Its antimicrobial activity against oral microorganisms such as Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus gordonii was examined, and its structural features were examined by CD and determined by two-dimensional (2D) nmr . The strains P . gingivalis (W50 and 381), A . actinomycetemcomitans (Y4 and 67), S . gordonii (DL1), and S . mutans (GS5) are found to be highly sensitive to this peptide at 2-2.5 microM concentrations, suggesting that the dodecapeptide is a potent antibiotic for oral pathogens . The weak negative n-sigma* band observed at approximately 265-270 nm in the CD spectra of this peptide provides evidence for the presence of a disulfide bridge . The negative n-pi* band at approximately 200 nm and the positive pi-pi* band at 185 nm suggest a folded structure for this peptide . The negative n-pi* shifts from 200 to 206 nm with an increase in intensity in dipalmitoylphosphotidylcholine vesicles, suggesting that the peptide might associate to form higher order aggregates in lipid medium . The assignment of backbone and side-chain proton resonances has been accomplished by the combined analysis of 2D total correlated and nuclear Overhauser effect spectroscopy . The temperature dependence of amide NH chemical shifts and (1)H-(2)H exchange effect on amide NH resonances indicate the involvement of amide NH groups of Cys3, Ile5, Ile8, Val10, and Arg12 in intramolecular hydrogen bonding . The coupling constant (J(NH-C(alpha)H)) values, the set of medium-, short-, and long-range nuclear Overhauser effects, and the results of restrained structure calculation using the distance geometry algorithm for nmr applications provide evidence for a folded, loop-like structure with a type I (III) beta-turn involving Ile5, Val6, Val7, and Ile8, and two antiparallel beta-strands involving the N-terminal Arg1, Leu2, Cys3, and Val4 and the C-terminal Arg9, Val10, Cys11, and Arg12 residues . The structure of the dodecapeptide mimics the amphiphilic structure of large 30-35 residue defensins and the peptide appears to exhibit similar antimicrobial potency . Hua Xi Yi Ke Da Xue Xue Bao, 1998 Jun, 29(2), 147 - 50 {Effects of surface roughness of two restorative materials on early Streptococcus sanguis adhesion in vitro}; Gao N et al.; This was an in vitro study aimed at the surface roughness of two kinds of commonly used restorative materials resin and alloy necessary to affect the adhesive behavior of Streptococcus sanguis(S . s) on them after 24 h . The surface roughness(SR) of each tested sample was evaluated with a profilometer quantitatively and observed with scanning electron microscope (SEM) morphologically . Then the adhesive microbial amount was determined by the clone forming unit counting method, and adhesion morphology was analyzed with SEM . The result showed a positive linear relation between the adhesion amount of S . s and the SR . The relative coefficients were rresin = 0.46(P < 0.01) and ralloy = 0.25(P < 0.01) respectively . These suggest the SR of the restorative material is important for the early adhesion of oral microbes in vitro . Therefore, before the prothesis is inserted in the patient's oral cavity, its surface should be polished as smoothly as possible so that the bacterial adhesive amount on its surface can be decreased, and hence the patient may keep in good oral health and have a prolonged use of the prothesis. Hua Xi Yi Ke Da Xue Xue Bao, 1997 Dec, 28(4), 353 - 6 {The immune response in rats immunized systemically by the surface protein antigen P1 from streptococcus mutans conjugated with procholeragenoid}; Luo Z et al.; This study was carried out to observe the antibody responses in rats after they were immunized with the surface protein P1 of streptococcus mutans when a special adjuvant was used . Antigen P1 was conjugated covalently with procholeragenoid (PCG), and then Sprague Dawley rats were immunized with P1 or the conjugated antigen P1-PCG subcutaneously or intragastrically . Anti-P1 antibody level was assayed at different time points by ELISA . The results showed that the levels of anti-P1 SIgA antibody in saliva rose when P1-PCG was given subcutaneously or intragastrically; the antibody level following the subcutaneous injection was higher and lasted longer, compared with that following the intragastric administretion . The level of anti-P1 IgG antibody in serum only rose when the rats were immunized subcutaneously . These results implied that mucosal immune response or humoral immune response could be induced subcutaneously when PCG was used as an adjuvant. Schweiz Med Wochenschr, 2000 Jan 22, 130(3), 72 - 6 {Spontaneous bacterial peritonitis with Streptococcus constellatus in an HIV positive patient}; Bielecki JW et al.; Liver diseases are an important cause of high morbidity and mortality in HIV-infected patients, and liver cirrhosis is the commonest cause of ascites in this population . We describe the case of a 38-year-old HIV-positive male (CDC stage B3, CD4 cell count 199/mm3) with a history of hepatitis C-associated liver cirrhosis . Following pneumonia he developed spontaneous bacterial peritonitis due to Streptococcus constellatus . Clinically noticeable was the gradually worsening course with few symptoms, despite the initially high ascitic fluid leucocyte count of over 11,000/microliter, but a favourable response to betalactam antibiotics. Proc Natl Acad Sci U S A, 2000 Feb 29, 97(5), 2235 - 40 Crystal structure of the zymogen form of the group A Streptococcus virulence factor SpeB: an integrin-binding cysteine protease; Kagawa TF et al.; Pathogenic bacteria secrete protein toxins that weaken or disable their host, and thereby act as virulence factors . We have determined the crystal structure of streptococcal pyrogenic exotoxin B (SpeB), a cysteine protease that is a major virulence factor of the human pathogen Streptococcus pyogenes and participates in invasive disease episodes, including necrotizing fasciitis . The structure, determined for the 40-kDa precursor form of SpeB at 1.6-A resolution, reveals that the protein is a distant homologue of the papain superfamily that includes the mammalian cathepsins B, K, L, and S . Despite negligible sequence identity, the protease portion has the canonical papain fold, albeit with major loop insertions and deletions . The catalytic site differs from most other cysteine proteases in that it lacks the Asn residue of the Cys-His-Asn triad . The prosegment has a unique fold and inactivation mechanism that involves displacement of the catalytically essential His residue by a loop inserted into the active site . The structure also reveals the surface location of an integrin-binding Arg-Gly-Asp (RGD) motif that is a feature unique to SpeB among cysteine proteases and is linked to the pathogenesis of the most invasive strains of S . pyogenes. Scand J Infect Dis, 1999, 31(6), 598 - 600 Streptococcus equinus endocarditis in a patient with pulmonary histiocytosis X; Sechi LA et al.; Although Streptococcus equinus has been isolated from the human bowel in an appreciable percentage of the adult general population, it has only rarely been described as a human pathogen . Our report describes the occurrence of S . equinus endocarditis in a patient who had no history of pre-existing heart disease, but who showed evidence of a late-stage pulmonary histiocytosis X . Endocarditis resolved promptly after antibiotic treatment, but required aortic valve substitution . Abnormalities of the immune system that have been demonstrated in patients with histiocytosis X could explain the occurrence of endocarditis in this patient. Scand J Infect Dis, 1999, 31(6), 597 - 8 Endocarditis caused by drug-resistant Streptococcus pneumoniae in a child; Pancharoen C et al.; We report a case of infective endocarditis caused by drug-resistant Streptococcus pneumoniae . Cefazolin or cefotaxime therapy induced a partial response . Treatment with vancomycin was successful . This microorganism may be more significant in endocarditis in areas with a high prevalence of drug-resistant Streptococcus pneumoniae. Indian J Med Res, 1999 Nov, 110, 164 - 8 Changing trend in susceptibility pattern of Streptococcus pneumoniae to penicillin in India; Lalitha MK et al.; Prior to 1995 all strains of Streptococcus pneumoniae isolated at a tertiary care hospital in south India were uniformly susceptible to penicillin . However, since late 1995 strains of S . pneumoniae with intermediate resistance to penicillin have been observed . Altogether there were 25 such isolates, 9 from invasive (5 from CSF as well as blood, 1 from pleural fluid and 3 from CSF alone) and 16 from noninvasive sites (6 from throat, 6 from sputum, 3 from eye and 1 from ear) respectively, thus 4.6 per cent of S . pneumoniae showed intermediate resistance of a total of 535 strains studied so far . The minimum inhibitory concentration (MIC) values of penicillin, erythromycin, chloramphenicol and cefotaxime were determined by agar dilution method and for confirmation, E test was carried out for penicillin alone . The MIC range obtained for penicillin was between 0.125-1.0 microgram/ml . Kirby-Bauer disc diffusion method was adopted for testing of erythromycin, chloramphenicol, co-trimoxazole, cefotaxime, tetracycline and vancomycin . We observed that none of the strains with intermediate resistance to penicillin were multidrug resistant . These strains belonged predominantly to serotype 14 (n = 10), 7B (n = 9), 19A (n = 3), 7F (n = 2) and 23F (n = 1) . Clonality was not observed in the 5 representative strains subjected to Box A finger printing method. Antimicrob Agents Chemother, 2000 Mar, 44(3), 798 - 801 A convenient assay for estimating the possible involvement of efflux of fluoroquinolones by Streptococcus pneumoniae and Staphylococcus aureus: evidence for diminished moxifloxacin, sparfloxacin, and trovafloxacin efflux; Beyer R et al.; We developed a simplified assay for estimating efflux by measuring the effect of reserpine on the growth of Streptococcus pneumoniae and Staphylococcus aureus over 7 h . Reserpine enhanced ciprofloxacin and levofloxacin 17 to 68% . The hydrophobic drug trovafloxacin and the drug moxifloxacin, with a bulky C-7 substituent but hydrophilicity similar to that of levofloxacin, showed little (0 to 11%) reserpine-enhancing effect . The ease of resistant mutant strain selection correlated with efflux susceptibility. Antimicrob Agents Chemother, 2000 Mar, 44(3), 790 - 3 Characterization of the Tn916-like transposon Tn3872 in a strain of abiotrophia defectiva (Streptococcus defectivus) causing sequential episodes of endocarditis in a child; Poyart C et al.; Clinical blood isolates from three sequential episodes of endocarditis occurring over a 6-month period in a child with a malformative cardiopathy were investigated . All isolates identified as Abiotrophia defectiva were resistant to erythromycin-clindamycin and to tetracycline-minocycline, due to the presence of sequences homologous to the erythromycin resistance gene ermB and to the tetracycline resistance gene tet(M), respectively . These resistance genes were located on a chromosomally borne composite Tn916-related transposon . These results demonstrate the involvement of conjugative transposons in the dissemination of antibiotic resistance in the genus Abiotrophia. Antimicrob Agents Chemother, 2000 Mar, 44(3), 787 - 9 Streptococcus gordonii strains resistant to fluorodeoxyuridine contain mutations in the thymidine kinase gene and are deficient in thymidine kinase activity; Franke CA et al.; Mutants of Streptococcus gordonii resistant to 5-fluorodeoxyuridine (FUdR(r)) were isolated . Each strain contained a point mutation resulting in the premature termination of the thymidine kinase (TK) open reading frame (tdk) . In vitro translation of the mutant tdk coding regions resulted in synthesis of truncated TK polypeptides deficient in TK activity. Antimicrob Agents Chemother, 2000 Mar, 44(3), 773 - 4 Comparative activities of ciprofloxacin and levofloxacin against Streptococcus pneumoniae in an In vitro dynamic model; Zinner SH et al.; The activities of levofloxacin (500 mg every 24 h) and ciprofloxacin (750 mg every 12 h) against six pneumococcal isolates in an in vitro dynamic model were compared . For one strain, levofloxacin reduced the inoculum by over 4 log CFU/ml and ciprofloxacin reduced the inoculum by over 2 log CFU/ml . For four isolates, both drugs reduced inocula by 4 log CFU/ml within 6 h, suggesting that this dose of ciprofloxacin should be as effective as levofloxacin against these pneumococci. Antimicrob Agents Chemother, 2000 Mar, 44(3), 767 - 70 Gemifloxacin is effective in experimental pneumococcal meningitis; Smirnov A et al.; In a rabbit model of Streptococcus pneumoniae meningitis, 5 mg of gemifloxacin mesylate (SB-265805) per kg/h reduced the bacterial titers in cerebrospinal fluid (CSF) almost as rapidly as 10 mg of ceftriaxone per kg/h (Deltalog CFU/ml/h +/- standard deviation {SD}, -0.25 +/- 0.09 versus -0.38 +/- 0.11; serum and CSF concentrations of gemifloxacin were 2.1 +/- 1.4 mg/liter and 0.59 +/- 0.38 mg/liter, respectively, at 24 h) . Coadministration of 1 mg of dexamethasone per kg did not affect gemifloxacin serum and CSF levels (2.7 +/- 1.4 mg/liter and 0.75 +/- 0.34 mg/liter, respectively, at 24 h) or activity (Deltalog CFU/ml/h +/- SD, -0.26 +/- 0.11). Antimicrob Agents Chemother, 2000 Mar, 44(3), 732 - 8 Mutations in ribosomal protein L16 conferring reduced susceptibility to evernimicin (SCH27899): implications for mechanism of action; Adrian PV et al.; A clinical isolate of Streptococcus pneumoniae (SP#5) that showed decreased susceptibility to evernimicin (MIC, 1.5 microgram/ml) was investigated . A 4,255-bp EcoRI fragment cloned from SP#5 was identified by its ability to transform evernimicin-susceptible S . pneumoniae R6 (MIC, 0.03 microgram/ml) such that the evernimicin MIC was 1.5 microgram/ml . Nucleotide sequence analysis of this fragment revealed that it contained portions of the S10-spc ribosomal protein operons . The nucleotide sequences of resistant and susceptible isolates were compared, and a point mutation (thymine to guanine) that causes an Ile52-Ser substitution in ribosomal protein L16 was identified . The role of this mutation in decreasing susceptibility to evernimicin was confirmed by direct transformation of the altered L16 gene . The presence of the L16 mutation in the resistant strain suggests that evernimicin is an inhibitor of protein synthesis . This was confirmed by inhibition studies using radiolabeled substrates, which showed that the addition of evernimicin at sub-MIC levels resulted in a rapid decrease in the incorporation of radiolabeled isoleucine in a susceptible isolate (SP#3) but was much less effective against SP#5 . The incorporation of isoleucine showed a linear response to the dose level of evernimicin . The incorporation of other classes of labeled substrates was unaffected or much delayed, indicating that these were secondary effects. Antimicrob Agents Chemother, 2000 Mar, 44(3), 598 - 601 Activities of trovafloxacin, gatifloxacin, clinafloxacin, sparfloxacin, levofloxacin, and ciprofloxacin against penicillin-resistant Streptococcus pneumoniae in an in vitro infection model; Hershberger E et al.; We adapted an in vitro pharmacodynamic model of infection to incorporate infected fibrin clots . The bactericidal activities of various fluoroquinolones against two strains of penicillin-resistant Streptococcus pneumoniae were studied over a 48-h period . Bacteria were prepared in Muller-Hinton broth by using colonies from a 24-h tryptic soy agar plus 5% sheep blood plate and were added to a mixture of cryoprecipitate (80%) and thrombin (10%) to achieve approxim