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| Curr Opin Investig Drugs, 2001 Jun, 2(6), 766 - 72 ABT-773 (Abbott Laboratories); Lawrence LE; ABT-773 is a macrolide antibacterial agent under development by Abbott Laboratories and Taisho Pharmaceutical Co Ltd for the potential treatment of bacterial infection {266579} . As of February 2001, ABT-773 had entered phase III trials in the US {398274} . Japanese phase II trials were expected to commence in June 2000 and a phase II trial is being designed for respiratory infections, with Abbott expecting filing in March 2002 {360455} . The bioavailability of ABT-773 in humans is unaffected by food {383228} and in a phase I, randomized, double-blind trial in healthy males only mild adverse effects, usually affecting the gastrointestinal system, were observed {383208} . Under an agreement, Abbott and Taisho are conducting joint research to discover new compounds; Abbott will have worldwide marketing, manufacturing and supply rights (except in Japan), and Taisho will receive royalties on Abbott's sales in consideration of granted rights . In Japan, the companies will co-market any resulting compounds {266579} . ABT-773 demonstrated good activity in vitro and in vivo against Streptococcus pneumoniae and Staphylococcus aureus {383229}, {383231}, and was highly potent even against macrolide-resistant {382149}, {382150} and invasive {383782} S pneumoniae. Curr Opin Investig Drugs, 2001 Jun, 2(6), 745 - 51 Antibacterial agents (oxazolidinones, quinolones, macrolides and new agents): patent highlights June to December 2000; Phillips OA et al.; Several patent disclosures were made on oxazolidinone, quinolone, macrolide and new antimicrobial agents between June and December 2000; 26 of these patents are discussed in this review . The exciting report on new oxazolidinone derivatives with potent activity against fastidious Gram-negative organisms is highlighted . Most of the new quinolones are structurally dissimilar to the traditional fluoroquinolones and demonstrate activity against Helicobacter pylori (MIC=0.013 microg/ml) . New 14-membered macrolides with broad-spectrum activity including against Mycobacterium avium are also presented . A novel peptidic antibiotic with potent antimicrobial activity against Streptococcus pneumoniae (MIC<0.0005 microg/ml) is also highlighted. Appl Environ Microbiol, 2001 Oct, 67(10), 4828 - 33 Streptococcus suis serotypes characterized by analysis of chaperonin 60 gene sequences; Brousseau R et al.; Streptococcus suis is an important pathogen of swine which occasionally infects humans as well . There are 35 serotypes known for this organism, and it would be desirable to develop rapid methods methods to identify and differentiate the strains of this species . To that effect, partial chaperonin 60 gene sequences were determined for the 35 serotype reference strains of S . suis . Analysis of a pairwise distance matrix showed that the distances ranged from 0 to 0.275 when values were calculated by the maximum-likelihood method . For five of the strains the distances from serotype 1 were greater than 0.1, and for two of these strains the distances were were more than 0.25, suggesting that they belong to a different species . Most of the nucleotide differences were silent; alignment of protein sequences showed that there were only 11 distinct sequences for the 35 strains under study . The chaperonin 60 gene phylogenetic tree was similar to the previously published tree based on 16S rRNA sequences, and it was also observed that strains with identical chaperonin 60 gene sequences tended to have identical 16S rRNA sequences . The chaperonin 60 gene sequences provided a higher level of discrimination between serotypes than the 16S RNA sequences provided and could form the basis for a diagnostic protocol. Arch Pathol Lab Med, 2001 Oct, 125(10), 1285 - 9 Method preferences and test accuracy of antimicrobial susceptibility testing: updates from the College of Amercian Pathologists Microbiology Surveys Program; Jones RN; College of American Pathologists Microbiology Resource Committe; OBJECTIVE: To summarize the antimicrobial susceptibility testing results from the College of American Pathologists (CAP) Microbiology Surveys Program for 2000 . Specifically, the frequency of tests used and the quantitative and qualitative (susceptibility category) accuracy were assessed . DESIGN: The CAP Microbiology Surveys challenged subscribers in 2000 with 3 well-characterized organisms for antimicrobial susceptibility testing in pure culture . Each laboratory was to use the test method and reporting procedures routinely applied to patient samples . The strains were National Committee for Clinical Laboratory Standards (NCCLS) quality control organisms with precisely defined antimicrobial susceptibility patterns and reproducibility . Results reported by participants (2685-2979/sample) were graded for categorical accuracy and quantitative performance by comparing reported minimal inhibitory concentrations (microg/mL) or zone diameters (mm) against quality control ranges published by the NCCLS . The appropriateness of reported drugs was determined in the context of the type and anatomic location of the infection . RESULTS: The tests most often used varied by the species of the organism and growth characteristics of the isolated strains . Nonfastidious, rapid-growing Surveys unknowns (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) were most often tested with commercial systems (MicroScan, 42.0%-42.4%; Vitek, 41.5%-43.0%) or with the standardized disk diffusion method (12.8%-13.9%) . In contrast, fastidious species, such as Streptococcus pneumoniae (ATCC 49619), were predominantly tested by Etest (40.3%), followed by disk diffusion (27.6%) and MicroScan (23.2%) . Categorical accuracy was essentially equal between dilution (98.9%) and diffusion (99.0%) methods . Among the minimal inhibitory concentration methods used to test penicillin against S pneumoniae, Etest method quantitative accuracy (96.3%) was greater than that of MicroScan (92.4%) . Quantitative accuracy was greatest for dilution minimal inhibitory concentration methods, with more than 90% of results within NCCLS quality control ranges for nearly all reported antimicrobials . Reevaluations of quality control ranges may be needed for 4 to 7 agents, depending on method . Reporting errors were also detected in 2 areas: (1) reporting results for drugs not active at the site of infection and (2) reporting results for drugs tested with suboptimal methods without published NCCLS interpretive criteria . CONCLUSIONS: Antimicrobial susceptibility testing methods used in US laboratories were dominated by commercial products with relatively high accuracy (qualitative and quantitative) . As available methods have become better suited to both fastidious and rapid-growing species, reporting errors have assumed a higher level of concern to the CAP Surveys in an effort to minimize prescription errors. Immunol Invest, 2001 Aug, 30(3), 191 - 207 A multi-laboratory evaluation of an enzyme-linked immunoassay quantitating human antibodies to Streptococcus pneumoniae polysaccharides; Quataert S et al.; An enzyme-linked immunoassay (EIA) is described and evaluated which quantitates human antibodies to serotype specific S . pneumoniae polysaccharide (PnPs) in human sera . Based on the observations previously described by Koskela (1), native PnPs are used as coating antigens and sera are absorbed with a soluble pneumococcal absorbant material containing C-polysaccharide (CPs) to ensure measurement of serotype specific anti-PnPs antibodies . The robustness of this method was evaluated by ten laboratories using the same reagents, protocol, and five human serum samples . Reproducible antibody values were obtained for IgM, IgG, and IgA antibodies to five different PnPs serotypes, 3, 6B, 14, 19F, and 23F . The overall mean percent coefficients of variation in this interlaboratory study for all five selotype specific anti-PnPs determinations with the five coded sera were 30% for IgG, 3/% for IgM, and 36% for IgA . This assay can be standardized for quantitation of serotype specific anti-PnPs antibodies, allowing comparison of antibody values in vaccine trials evaluating pneumococcal vaccines. Mil Med, 2001 Sep, 166(9), 759 - 63 Prospective study of respiratory infections at the U.S . Naval Academy; Gray GC et al.; Midshipmen at the U.S . Naval Academy have recently suffered epidemics of upper respiratory tract infections . Seeking to determine cause, in June 1998 we enrolled 1,243 (99.5%) of 1,249 new midshipmen (plebes) and followed them during their first 11 months of training . Eighty-five plebes sought medical attention for acute respiratory disease . Using culture, serologic studies, and polymerase chain reaction, considerable evidence for respiratory pathogen infection was found among the ill subjects: Chlamydia pneumoniae in 41 (52.6%), Mycoplasma pneumoniae in 19 (25.3%), influenza in 11 (14.2%), Streptococcus pneumoniae in 6 (7.3%), and adenovirus in 1 (1.2%) . Additionally, 873 (81%) the 1,077 plebes who completed an end-of-year questionnaire complained of having one or more respiratory symptoms (> 12 hours) during their first year of school . Of these, 132 (15%) reported that the symptoms significantly affected their performance . Study results suggest that respiratory infections were frequent, had a significant adverse impact on training, and were often attributable to bacterial pathogens. Am J Obstet Gynecol, 2001 Sep, 185(3), 586 - 92 An in vitro study of antibacterial properties of the cervical mucus plug in pregnancy; Hein M et al.; OBJECTIVE: To evaluate whether cervical mucus plugs are antibacterial in vitro . STUDY DESIGN: Cervical mucus plugs from 56 healthy women in labor were studied by 2 different antimicrobial assays: (1) analysis of the inhibition by the cervical mucus plug of several gram-positive and gram-negative bacteria by overlaying the cervical mucus plug onto an agar plate with imbedded bacteria, and (2) determination of the antibacterial property of the cervical mucus plug material by radial diffusion assay with group B Streptococcus and Escherichia coli . RESULTS: In the agar overlay assay, there was complete inhibition of clinical isolates of Staphylococcus saprophyticus, E coli, and Pseudomonas aeruginosa and patient-variable partial-to-complete inhibition of Enterococcus faecium, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus agalactiae . In the radial diffusion assay, cervical mucus plugs had activity toward group B Streptococcus equivalent to 0.075 microg/mL of gentamicin and toward E coli equivalent to 0.5 microg/mL of gentamicin . CONCLUSION: A low-molecular substance with antibacterial activity in the cervical mucus plug may protect the fetus against ascending infections. Dent Mater, 2001 Nov, 17(6), 485 - 91 Inhibitory effects of resin composite containing bactericide-immobilized filler on plaque accumulation; Ebi N et al.; OBJECTIVE: Previously, we have reported that incorporation of the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) was effective in immobilizing bactericide in the resin matrix, and an antibacterial composite without release of the agent could be achieved . In this study, an attempt was made to increase the density of bactericide immobilized in composite, and the inhibitory effects of this modified antibacterial composite on plaque accumulation were determined, focusing on the reliability of the effects and the mechanisms to affect the plaque formation . METHODS: An experimental composite containing immobilized bactericide at 2.83% was prepared by the incorporation of MDPB into a prepolymerized resin filler of control composite, and elution of antibacterial components and inhibition of in vitro plaque accumulation by Streptococcus mutans were determined . The inhibitory effects of the experimental composite on the attachment, glucan synthesis and growth of S . mutans on the surface were also examined in addition to the comparison of surface roughness and hydrophobicity with controls . The results were analyzed using the Student's t-test . RESULTS: The experimental composite had reproducible inhibitory effects against plaque accumulation compared with control (p<0.05), although it showed no elution of unpolymerized MDPB . The plaque-inhibitory effect of the experimental composite was found to depend upon the ability to inhibit the attachment, glucan synthesis, and growth of bacteria on its surface as no significant differences in the surface characteristics were obtained between control and experimental composites (p>0.05) . SIGNIFICANCE: It was indicated that the experimental composite containing bactericide-immobilized filler has the possibility to be used clinically with an effective anti-plaque property. J Bacteriol, 2001 Oct, 183(20), 5964 - 73 uvrA is an acid-inducible gene involved in the adaptive response to low pH in Streptococcus mutans; Hanna MN et al.; The pH-inducible acid tolerance response (ATR) is believed to play a major role in acid adaptation and virulence of Streptococcus mutans . To study this phenomenon in S . mutans JH1005, differential display PCR was used to identify and clone 13 cDNA products that had increased expression in response to pH 5.0 compared to that of pH 7.5-grown cells . One of these products, confirmed to be pH inducible by RNA dot blot and reverse transcription-PCR analyses, had 67% identity to a uvrA-UV repair excinuclease gene in Bacillus subtilis . Further sequence analysis of the uvrA homologue using the S . mutans genome database revealed that the complete gene was encoded in an open reading frame (ORF) of 2,829 bp (944 amino acids; 104.67 kDa) . Immediately 3' of uvrA was an ORF encoding a putative aminopeptidase gene (pepP) . uvrA knockouts were constructed in S . mutans strains JH1005, NG8, and UA159 using allelic-exchange mutagenesis, replacing the entire gene with an erythromycin resistance cassette . As with uvrA mutants in other bacteria, the S . mutans uvrA mutants were extremely sensitive to UV irradiation . The uvrA mutant of S . mutans JH1005 was also more sensitive than the wild type to growth at pH 5.0, showing a 15% reduction in growth rate and a 14% reduction in final resting culture density . Acid-adapted S . mutans JH1005 uvrA mutants were shown to be more resistant to UV irradiation than was the parent but were unable to survive exposure to a killing pH of 3.0 . Moreover, agarose gel electrophoretic analysis of chromosomal DNA isolated from uvrA-deficient cells exposed to low pH demonstrated more DNA damage than that for the wild-type strain . Here we suggest that uvrA and the nucleotide excision repair pathway are involved in the repair of acid-induced DNA damage and are associated with successful adaptation of S . mutans to low pH. Arch Cardiol Mex, 2001 Apr-Jun, 71(2), 127 - 35 {Rheumatic fever in the 5-year period of 1994-1999 at 2 hospitals in San Luis Potosi and Mexico D.F.}; Soto Lopez ME et al.; OBJECTIVE: To assess the incidence of acute Rheumatic Fever (ARF) . PATIENTS AND METHODS: A retrospective, descriptive, observational study on the first attack and recurrence was performed in a general hospital and a reference center . RESULTS: By Jones criteria: 67 cases, 39 women and 28 men; 58% first attack, 42% recurrence . Higher incidence during spring-winter . The most common major criteria were: carditis, polyarthritis . The most common minor criteria were: fever, arthralgias and acute phase reaction markers . No differences between hospitals were noted . Evidence of contact with streptococcus was found . Mitral, aortic and tricuspid valves were commonly affected . Incidence in the age group > 5 < 20 was 7/1000 . DISCUSSION: Incidence of ARF has decreased, but has not been eradicated . It occurs in developing countries, where it remains an issue of public health . Failures in clinical suspicion, prophylaxis, and adherence to treatment influence this situation . Education for health, early diagnosis, and primary and secondary prophylaxis should be reinforced. Clin Infect Dis, 2001 Oct 15, 33(8), 1324 - 8 Epub 2001 Sep 17. Time to positivity of blood cultures for children with Streptococcus pneumoniae bacteremia; Neuman MI et al.; We report on a 3-year (1 January 1996 through 31 December 1999) retrospective chart review of children with Streptococcus pneumoniae bacteremia to identify the time to identification of growth of S . pneumoniae in blood culture and to attempt to identify clinical predictors of early versus late growth of S . pneumoniae in culture . The time to detection of S . pneumoniae in blood culture for immunocompetent patients ranged from 4.4 to 25.9 hours (h), with a mean of 11.5 h (standard deviation, 2.8) . There was no difference in the time to detection for immunocompromised versus immunocompetent patients . The 10th and 90th deciles for time to detection among immunocompetent patients were 9.2 and 14.0 h, respectively . There were no differences in white blood cell count, absolute neutrophil count, or height of fever between the lowest and highest decile groups . Ninety percent of blood cultures yielding S . pneumoniae are noted positive within 14 h, and no clinical or laboratory parameters accurately predicted early versus late growth of S . pneumoniae in blood culture. Semin Respir Infect, 2001 Sep, 16(3), 186 - 95 Drug-resistant Streptococcus pneumoniae infections: clinical importance, drug treatment, and prevention; Kellner JD; Drug-resistant Streptococcus pneumoniae (DRSP) have complicated the management of pneumococcal infections throughout the world in recent years . Important risk factors for the development of a DRSP infection include recent antibiotic use, young age, immunosuppression, daycare center attendance, and recent hospitalizations . Although intermediate resistance to beta-lactam antibodies does not appear to have an impact on the mortality of S . pneumoniae infections, evidence is accumulating to suggest that more highly resistant isolates may increase mortality . Clinical and bacteriologic failures are more common in DRSP meningitis and otitis media . To select appropriate therapy, one must consider whether a patient has risk factors for resistance, the site and severity of the infection, and antibiotic susceptibility data . Reducing the impact of DRSP may be achieved through reducing antibiotic use in communities and increased understanding of other factors that contribute to the development and transmission of resistance . The most important way to reduce all S . pneumoniae infections is to increase the use of existing polysaccharide vaccines and to begin to use new polysaccharide-protein conjugate vaccines in young children . Semin Respir Infect, 2001 Sep, 16(3), 177 - 85 Antimicrobial resistance with Streptococcus pneumoniae: much ado about nothing? Doern GV. During the past decade in the United States, Streptococcus pneumoniae has changed dramatically in the context of antimicrobial resistance . Resistance to multiple different antibiotic classes including beta-lactams (penicillins, cephalosporins, and beta-lactamase inhibitor combinations), macrolides, clindamycin, the tetracyclines, chloramphenicol, and trimethoprim-sulfamethoxazole (TMP/SMX) has emerged at high rates with this important respiratory tract pathogen . There is no question that the in vitro activity of these antimicrobial agents is substantially lower for many strains of S . pneumoniae than it was even a few years ago . The larger question is, however, what does this decrease in activity mean from a clinical perspective? Stated another way, does resistance defined according to current standards in the laboratory, translate into diminished effectiveness when these agents are used to treat patients with pneumococcal infections? It is this question that serves as the principal basis for this review . Med Clin (Barc), 2001 Sep 15, 117(7), 241 - 5 {Pneumococcal bacteremia in patients aged over 65 years . A study of 161 cases}; Vaqueiro Subirats M et al.; BACKGROUND: Here we present the results from a prospective analysis of pneumococcal bacteremia in patients older than 65 years, highlighting its differences with regard to patients aged from 15 to 64 years . PATIENTS AND METHOD: We performed a prospective follow-up study (1988-1999) of patients older than 14 years whohad blood cultures positive for Streptococcus pneumoniae(S . pneumoniae) . S . pneumoniae was identified using standard techniques . The antimicrobial sensitivity was analysed using the broth midrodilution technique and the serotype was studied in a reference laboratory . Statistical analyses were performed by means of the SPSS program, version 9.0 . RESULTS: 161 out of 321 bacteremias occurred in 154 patients older than 65 years (86 males) . Most frequent underlying diseases were chronic obstructive pulmonary disease (COPD), diabetes and neoplasia . Most patients had pneumonia, 29.7% with ilfiltrates involving at least two lung lobes . 9.4% of bacteremias were nosocomial . Among patients older than 65 years, 32.5% of strains were resistant to penicillin, compared to 21.7% in patients younger than 65 years(p = 0.034) . Mortality in patients over 65 years was higher than in younger patients (25.3% vs 11.5%; p = 0.002) and mortality predictive factors in a multivariate analysis were age, nosocomial bacteremia and multi-lobe involvement (in patients with pneumonia) . CONCLUSIONS: There is a high incidence of pneumococcal bacteremia in patients over 65 years of age . These patients have a higher rate of resistance to penicillin and a higher mortality rate than younger patients.Mortality predictive factors are age, nosocomial bacteremia and multi-lobe involvement. Int Immunopharmacol, 2001 Sep, 1(9-10), 1789 - 95 Severe impairment of anti-cancer effect of lipoteichoic acid-related molecule isolated from a penicillin-killed Streptococcus pyogenes in toll-like receptor 4-deficient mice; Okamoto M et al.; A lipoteichoic acid-related molecule (OK-PSA) isolated from OK-432, a penicillin-killed Streptococcus pyogenes, is a potent inducer of Th1 cytokines, and elicits anti-cancer effect in tumor-bearing mice . Toll-like receptor (TLR) 4 is a member of the recently identified toll-like receptor family of proteins that has been implicated in lipopolysaccharide-induced cell signaling . In the present study, we have examined the role of TLR4 for OK-PSA-induced Th1-cytokine production and anti-tumor effect by using C3H/HeJ mice in which TLR4 function is impaired . Although OK-PSA strikingly induced Th1 cytokines {interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, IL-12 and IL-18} in the splenocytes derived from control animals (C3H/HeN), OK-PSA did not induce the cytokines in the splenocytes from C3H/HeJ . Furthermore, C3H/HeJ-derived splenocytes acquired the responsiveness to OK-PSA stimulation by overexpression of TLR4 gene . Finally, OK-PSA administration significantly inhibited the tumor growth and lung metastasis of syngeneic squamous cell carcinoma cells in C3H/HeN; however, no effect of OK-PSA was observed in C3H/HeJ . These findings strongly suggest that TLR4 signaling is involved in regulating OK-PSA-induced anti-cancer immunity. Epidemiol Infect, 2001 Aug, 127(1), 49 - 55 Prevalence of carriage of antimicrobial resistant strains of Streptococcus pneumoniae in primary school children in Hong Kong; Boost MV et al.; A cross-sectional survey was conducted to determine prevalence and assess risk factors for carriage of antibiotic resistant strains of Streptococcus pneumoniae in healthy school children in Hong Kong . Throat swabs were collected from 1455 subjects and written questionnaires providing demographic data and medical history were completed by parents . The overall carriage rate of Streptococcus pneumoniae was 3.5%, of which 49% were penicillin resistant . High levels of resistance to tetracycline (73%), erythromycin (52%), trimethoprim (66%) and ciprofloxacin (57%) were observed . Carriage was associated with presence of a younger sibling (OR = 1.79) and use of antibiotics (OR = 2.31) . High use of day care and small size of housing units did not result in a high rate of carriage . The low rate of carriage may be linked to high use of antibiotics, geographical factors or ethnicity . High rates of antibiotic resistance reflect heavy use of antibiotics by general practitioners. Eur J Clin Microbiol Infect Dis, 2001 Jul, 20(7), 494 - 7 Relative potency of telithromycin, azithromycin and erythromycin against recent clinical isolates of gram-positive cocci; Barry AL et al.; A ketolide (telithromycin), an azalide (azithromycin) and a macrolide (erythromycin) were tested against 2,733 isolates of gram-positive cocci gathered from 11 different medical centers . Telithromycin was active against erythromycin-resistant staphylococci that were susceptible to clindamycin but was not active against those that were resistant to clindamycin . More than 99% of all Streptococcus pneumoniae and Streptococcus pyogenes isolates were susceptible to 1 microg/ml of telithromycin including erythromycin- and azithromycin-resistant strains . Telithromycin was not only more potent than azithromycin against macrolide-susceptible strains, it was also active against most macrolide-resistant strains . Although the prevalence of macrolide-resistant pneumococci increased from 19% to 27% between 1997 and 1999, macrolide resistance among other gram-positive cocci did not change substantially in that 2-year period. Eur J Clin Microbiol Infect Dis, 2001 Jul, 20(7), 490 - 3 Reduced release of DNA from streptococcus pneumoniae after treatment with rifampin in comparison to spontaneous growth and ceftriaxone treatment; Gerber J et al.; In order to study the release of DNA from Streptococcus pneumoniae in vitro during spontaneous growth and treatment with ceftriaxone or rifampin, a semiquantitative polymerase chain reaction was used . During spontaneous growth, high concentrations of bacterial DNA were released . Exposure to 10 microg/ml of ceftriaxone decreased the DNA release, in median, by 19 times (P=0.03 vs . spontaneous growth) . Treatment with 10 microg/ml of rifampin led to a reduction of DNA release, in median, by a factor of 49 (P=0.03 vs . ceftriaxone; six experiments performed on different days). Chemotherapy, 2001 Sep-Oct, 47(5), 354 - 8 In vitro activity of novel fluoroquinolones against Streptococcus pneumoniae isolated from children with acute otitis media; Yagupsky P et al.; BACKGROUND: In recent years, novel fluoroquinolones with improved activity against gram-positive organisms have been introduced into clinical practice . These drugs may be of potential benefit for the treatment of pneumococcal otitis media, including infections caused by organisms resistant to conventional drugs . METHODS: In vitro activity of 6 fluoroquinolones against 77 pneumococcal isolates from middle-ear fluid was determined by the E test . RESULTS: Resistance to penicillin, co-trimoxazole, erythromycin, clindamycin, and tetracycline was present in 59 (76.6%), 47 (61.0%), 19 (24.7%), 11 (14.3%), and 17 (22.1%) isolates, respectively . Fluoroquinolone MIC(50) and MIC(90) (in microg/ml) were as follows: ciprofloxacin: 1.0 and 3.0, levofloxacin: 0.75 and 1.0, sparfloxacin: 0.25 and 0.38, grepafloxacin: 0.25 and 0.38, trovafloxacin: 0.094 and 0.125, and moxifloxacin: 0.19 and 0.25, respectively . CONCLUSIONS: Novel fluoroquinolones and especially trovafloxacin and moxifloxacin appear to be of potential value for the treatment of acute otitis media caused by pneumococci resistant to traditional antibiotics . Jpn J Antibiot, 2001 Jul, 54(7), 365 - 71 {In vitro and in vivo activities of panipenem against penicillin-resistant Streptococcus pneumoniae}; Fukuoka T et al.; Efficacy of panipenem/betamipron (PAPM/BP) against experimental pneumonia caused by penicillin-resistant Streptococcus pneumoniae (PRSP: MIC of benzylpenicillin, > or = 1.56 micrograms/ml) in mice was compared with those of imipenem/cilastatin (IPM/CS), meropenem (MEPM), cefozopran (CZOP), ceftriaxone (CTRX), ampicillin (ABPC), and vancomycin (VCM) . The infection was induced by inoculating a PRSP clinical isolate, 9601 (serotype 6) or 10,693 (serotype 19), into ddY male mice intranasally . Drugs were administered subcutaneously at doses of 0.4, 2, and 10 mg/kg, 18, 26, 42, and 50 hours post-infection . Viable cell counts in the lungs were determined 66 hours post-infection . PAPM/BP showed the greatest efficacy against the infections among tested drugs . MICs of PAPM against PRSP 9601 and 10,693 were both 0.125 microgram/ml, which were superior to those of IPM (0.25 and 0.5 microgram/ml, respectively), MEPM (0.5 and 1 microgram/ml, respectively), CZOP (2 and 1 microgram/ml, respectively), CTRX (both 1 microgram/ml), ABPC (both 4 micrograms/ml), and VCM (0.5 and 0.25 microgram/ml, respectively) . These results suggest that the potent in vivo activity of PAPM/BP reflects the potent in vitro activity of PAPM . MICs of PAPM, IPM, MEPM, and CZOP against clinical isolates, penicillin-susceptible S . pneumoniae (PSSP: MIC of benzylpenicillin, < or = 0.05 microgram/ml), penicillin-intermediate S . pneumoniae (PISP: MIC of benzylpenicillin, 0.1-0.78 microgram/ml), and PRSP, were tested by an agar dilution method . MIC90s of the drugs against the PSSP, PISP, and PRSP were as follows: PAPM, 0.012, 0.05, and 0.39 microgram/ml; IPM, < or = 0.006, 0.1, and 0.78 microgram/ml; MEPM, 0.05, 0.39, and 1.56 micrograms/ml; and CZOP, 0.2, 0.78, and 6.25 micrograms/ml, respectively . Thus, PAPM showed the most potent activity among tested drugs against clinical isolates of PISP and PRSP. Antimicrob Agents Chemother, 2001 Oct, 45(10), 2955 - 7 Streptococcus pneumoniae isolates with reduced susceptibility to ciprofloxacin in Spain: clonal diversity and appearance of ciprofloxacin-resistant epidemic clones; Alou L et al.; Analysis of the pulsed-field gel electrophoretic profiles of 82 pneumococcal isolates with reduced susceptibility to ciprofloxacin (RSC) and of 90 co-occurring susceptible isolates indicates a considerable genetic diversity among isolates with RCS and points to a close relation between the two groups . This finding suggests that pneumococci with RCS emerge through independent mutational events. Antimicrob Agents Chemother, 2001 Oct, 45(10), 2916 - 21 Pharmacodynamics of gemifloxacin against Streptococcus pneumoniae in an in vitro pharmacokinetic model of infection; MacGowan AP et al.; The pharmacodynamics of gemifloxacin against Streptococcus pneumoniae were investigated in a dilutional pharmacodynamic model of infection . Dose fractionation was used to simulate concentrations of gemifloxacin in human serum associated with 640 mg every 48 h (one dose), 320 mg every 24 h (two doses), and 160 mg every 12 h (four doses) . Five strains of S . pneumoniae for which MICs were 0.016, 0.06, 0.1, 0.16, and 0.24 mg/liter were used to assess the antibacterial effect of gemifloxacin . An inoculum of 10(7) to 10(8) CFU/ml was used, and each experiment was performed at least in triplicate . The pharmacodynamic parameters (area under the concentration-time curve {AUC}/MIC, maximum concentration of drug in serum {C(max)}/MIC, and the time that the serum drug concentration remains higher than the MIC {T > MIC}) were related to antibacterial effect as measured by the area under the bacterial-kill curve from 0 to 48 h (AUBKC(48)) using an inhibitory sigmoid E(max) model . Weighted least-squares regression was used to predict the effect of the pharmacodynamic parameters on AUBKC(48), and Cox proportional-hazards regression was used to predict the effect of the three pharmacodynamic parameters on the time needed to kill 99.9% of the starting inoculum (T99.9) . There was a clear relationship between strain susceptibility and clearance from the model . The simulations (160 mg every 12 h) were associated with slower initial clearance than were the other simulations; in contrast, bacterial regrowth occurred with the 640-mg simulation when MICs were > or =0.1 mg/liter . The percentage coefficient of variance was 19% for AUBKC(48), and the inhibitory sigmoid E(max) model best fit the relationship between AUBKC(48) and AUC/MIC . C(max)/MIC and T > MIC fit less well . The maximum response occurred at an AUC/MIC of >300 to 400 . In weighted least-squares regression analysis, there was no evidence that C(max)/MIC was predictive of AUBKC(48), but both AUC/MIC and T > MIC were . A repeat analysis using only data for which the T > MIC was >75% and for which hence regrowth was minimized indicated that AUC/MIC alone was predictive of AUBKC(48) . Initial univariate analysis indicated that all three pharmacodynamic parameters were predictive of T99.9, but in the multivariate model only C(max)/MIC reached significance . These data indicate that gemifloxacin is an effective antipneumococcal agent and that AUC/MIC is the best predictor of antibacterial effect as measured by AUBKC(48) . However, C(max)/MIC is the best predictor of speed of kill, as measured by T99.9 . T > MIC also has a role in determining AUBKC(48), especially when the dose spacing is considerable . Once-daily dosing seems most suitable for gemifloxacin. Antimicrob Agents Chemother, 2001 Oct, 45(10), 2865 - 70 Selection and genetic characterization of Streptococcus pneumoniae mutants resistant to the des-F(6) quinolone BMS-284756; Hartman-Neumann S et al.; Existing quinolones are known to target the type II topoisomerases in bacteria . In order to determine which of these targets are of key importance in Streptococcus pneumoniae treated with BMS-284756 (T-3811ME), a novel des-F(6) quinolone, resistant mutants were selected in several steps of increasing resistance by plating pneumococci on a series of blood agar plates containing serial twofold-increasing concentrations of drug . After incubation, colonies that arose were selected and passaged twice on antibiotic-containing media at the selection level . Mutants generally showed increases in resistance of four- to eightfold over the prior level of susceptibility . Mutants in the next-higher level of resistance were selected from the previous round of resistant mutants . Subsequently, chromosomal DNA was prepared from parental (R6) pneumococci and from at least three clones from each of four levels of increasing antibiotic resistance . Using PCR primers, 500- to 700-bp amplicons surrounding the quinolone resistance determining regions (QRDR) of gyrA, gyrB, parC, and parE genes were prepared from each strain . Internal primers were used to sequence both DNA strands in the regions of approximately 400 bp centered on the QRDR . Mutations identified with increasing levels of resistance included changes in GyrA at Ser-81 and Glu-85 and changes in ParC at Ser-79 and Asp-83 . Changes in GyrB and ParE were not observed at the levels of resistance obtained in this selection . The resistance to comparator quinolones (levofloxacin, ciprofloxacin, and moxifloxacin) also increased in four- to eightfold steps with these mutations . The intrinsically greater level of antibacterial activity and thus lower MICs of BMS-284756 observed at all resistance levels in this study may translate to coverage of these resistant pneumococcal strains in the clinic. Antimicrob Agents Chemother, 2001 Oct, 45(10), 2793 - 7 Pharmacodynamics of fluoroquinolones against Streptococcus pneumoniae in patients with community-acquired respiratory tract infections; Ambrose PG et al.; Fluoroquinolone antibiotic agents have demonstrated efficacy in the treatment of respiratory tract infections . This analysis was designed to examine the relationship between drug exposure, as measured by the free-drug area under the concentration-time curve at 24 h (AUC(24))/MIC ratio, and clinical and microbiological responses in patients with community-acquired respiratory tract infections involving Streptococcus pneumoniae . The study population included 58 adult patients (34 males, 24 females) who were enrolled in either of two phase III, randomized, multicenter, double-blind studies of levofloxacin versus gatifloxacin for the treatment of community-acquired pneumonia or acute exacerbation of chronic bronchitis . Clearance equations from previously published population pharmacokinetic models were used in conjunction with dose and adjusted for protein binding to estimate individual patient free-drug AUC(24)s . In vitro susceptibility was determined in a central laboratory by broth microdilution in accordance with NCCLS guidelines . Pharmacodynamic analyses were performed on data from all evaluable patients with documented S . pneumoniae infection using univariate and multivariable logistic regression; pharmacodynamic breakpoints were estimated using Classification and Regression Tree analysis . A statistically significant (P = 0.013) relationship between microbiological response and the free-drug AUC(24)/MIC ratio was detected . At a free-drug AUC(24)/MIC ratio of <33.7, the probability of a microbiological response was 64%, and at a free-drug AUC(24)/MIC ratio of >33.7, it was 100% (P < 0.01) . These findings may provide a minimum target free-drug AUC(24)/MIC ratio for the treatment of infections involving S . pneumoniae with fluoroquinolone antibiotics and provide a paradigm for the selection of fluoroquinolones to be brought forward from drug discovery into clinical development and dose selection for clinical trials . Further, when target free-drug AUC(24)/MIC ratios are used in conjunction with stochastic modeling techniques, these findings may be used to support susceptibility breakpoints for fluoroquinolone antibiotics and S . pneumoniae. Antimicrob Agents Chemother, 2001 Oct, 45(10), 2765 - 70 Nasopharyngeal carriage of antimicrobial-resistant Streptococcus pneumoniae among young children attending 79 kindergartens and day care centers in Hong Kong; Chiu SS et al.; Resistance to penicillin and multiple antimicrobial agents among Streptococcus pneumoniae strains is becoming an increasing problem worldwide and in Asia . To determine the prevalence of carriage of S . pneumoniae isolates not susceptible to penicillin in young children, we obtained nasopharyngeal swab specimens from 1,978 children (ages, 2 to 6 years) attending 79 day care centers or kindergartens . Three hundred eighty-three strains of S . pneumoniae were isolated from these children . Fifty-eight percent of these isolates had reduced susceptibility to penicillin, 123 (32.1%) were intermediate, and 100 (26.1%) were resistant . A very high penicillin MIC (4 microg/ml) was found in 3.3% of the isolates . The isolates also demonstrated high rates of resistance to other antimicrobial agents (51.2% to cefaclor, 50.2% to cefuroxime, 42.8% to cefotaxime, 80.7% to trimethoprim-sulfamethoxazole, 77% to erythromycin, 60% to clindamycin, and 33.7% to chloramphenicol) . No isolate was resistant to fluoroquinolone . Multidrug resistance (not susceptible to the beta-lactams and three or more other classes) was found in 39.4% of the isolates . Risk factors for the carriage of S . pneumoniae not susceptible to penicillin were multiple physician visits in the preceding 3 months and use of antimicrobial agents by the individual or by household members in the preceding 3 months . In the logistic regression analysis, only the use of antimicrobial agents in the preceding 3 months was an independent risk factor (P = 0.004; odds ratio, 2; 95% confidence interval, 1.2 to 3.2) . This study demonstrated the high prevalence of antibiotic-resistant S . pneumoniae in healthy young children in the community in Hong Kong. Free Radic Biol Med, 2001 Sep 15, 31(6), 754 - 62 Oxidative stress in brain during experimental bacterial meningitis: differential effects of alpha-phenyl-tert-butyl nitrone and N-acetylcysteine treatment; Christen S et al.; Antioxidant treatment has previously been shown to be neuroprotective in experimental bacterial meningitis . To obtain quantitative evidence for oxidative stress in this disease, we measured the major brain antioxidants ascorbate and reduced glutathione, and the lipid peroxidation endproduct malondialdehyde in the cortex of infant rats infected with Streptococcus pneumoniae . Cortical levels of the two antioxidants were markedly decreased 22 h after infection, when animals were severely ill . Total pyridine nucleotide levels in the cortex were unaltered, suggesting that the loss of the two antioxidants was not due to cell necrosis . Bacterial meningitis was accompanied by a moderate, significant increase in cortical malondialdehyde . While treatment with either of the antioxidants alpha-phenyl-tert-butyl nitrone or N-acetylcysteine significantly inhibited this increase, only the former attenuated the loss of endogenous antioxidants . Cerebrospinal fluid bacterial titer, nitrite and nitrate levels, and myeloperoxidase activity at 18 h after infection were unaffected by antioxidant treatment, suggesting that they acted by mechanisms other than modulation of inflammation . The results demonstrate that bacterial meningitis is accompanied by oxidative stress in the brain parenchyma . Furthermore, increased cortical lipid peroxidation does not appear to be the result of parenchymal oxidative stress, because it was prevented by NAC, which had no effect on the loss of brain antioxidants. Oral Microbiol Immunol, 2001 Oct, 16(5), 319 - 20 Actinomyces naeslundii fimbrial protein Orf977 shows similarity to a streptococcal adhesin; Hoflack L et al.; The nucleotide sequence of the chromosomal DNA, upstream of Actinomyces naeslundii T14V fimbrial gene fimA, was determined . One open reading frame (orf977) encoding 977 amino acids was found, preceded by a gene homologous to elongation factor TU . Database searches revealed that Orf977 was homologous to CshA, a Streptococcus gordonii protein involved in cell adhesion . Previous studies had already determined two genes in the type 2 fimbrial gene cluster of A . naeslundii T14V: the structural subunit fimA, and orf365 with unknown function, followed by ribosomal genes . This study completes the type 2 fimbrial gene cluster sequence. J Biol Chem, 2001 Nov 30, 276(48), 45106 - 12 Epub 2001 Sep 11. Crystal structure of PBP2x from a highly penicillin-resistant Streptococcus pneumoniae clinical isolate: a mosaic framework containing 83 mutations; Dessen A et al.; Penicillin-binding proteins (PBPs) are the main targets for beta-lactam antibiotics, such as penicillins and cephalosporins, in a wide range of bacterial species . In some Gram-positive strains, the surge of resistance to treatment with beta-lactams is primarily the result of the proliferation of mosaic PBP-encoding genes, which encode novel proteins by recombination . PBP2x is a primary resistance determinant in Streptococcus pneumoniae, and its modification is an essential step in the development of high level beta-lactam resistance . To understand such a resistance mechanism at an atomic level, we have solved the x-ray crystal structure of PBP2x from a highly penicillin-resistant clinical isolate of S . pneumoniae, Sp328, which harbors 83 mutations in the soluble region . In the proximity of the Sp328 PBP2x* active site, the Thr(338) --> Ala mutation weakens the local hydrogen bonding network, thus abrogating the stabilization of a crucial buried water molecule . In addition, the Ser(389) --> Leu and Asn(514) --> His mutations produce a destabilizing effect that generates an "open" active site . It has been suggested that peptidoglycan substrates for beta-lactam-resistant PBPs contain a large amount of abnormal, branched peptides, whereas sensitive strains tend to catalyze cross-linking of linear forms . Thus, in vivo, an "open" active site could facilitate the recognition of distinct, branched physiological substrates. J Biol Chem, 2001 Nov 30, 276(48), 44551 - 6 Epub 2001 Sep 11. Substrate specificity of the streptococcal cysteine protease; Nomizu M et al.; The streptococcal pyrogenic exotoxin B (SpeB) is an important factor in mediating Streptococcus pyogenes infections . SpeB is the zymogen of the streptococcal cysteine protease (SCP), of which relatively little is known regarding substrate specificity . To investigate this aspect of SCP function, a series of internally quenched fluorescent substrates was designed based on the cleavage sites identified in the autocatalytic processing of SpeB to mature SCP . The best substrates for SCP contain three amino acids in the nonprimed position (i.e . AIK in P(3)-P(2)-P(1)) . Varying the length of the substrate on the primed side of the scissile bond has a relatively lower effect on activity . The highest activity (k(cat)/K(M) = 2.8 +/- 0.6 (10(5) x m(-1)s(-1)) is observed for the pentamer 3-aminobenzoic acid-AIKAG-3-nitrotyrosine, which spans subsites S(3) to S(2)' on the enzyme . High pressure liquid chromatography and mass spectrometry analyses show that the substrates are cleaved at the site predicted from the autoprocessing experiments . These results show that SCP can display an important level of endopeptidase activity . Substitutions at position P(2) of the substrate clearly indicate that the S(2) subsite of SCP can readily accommodate substrates containing a hydrophobic residue at that position and that some topological preference exists for that subsite . Substitutions in positions P(3), P(1), and P(1)' had little or no effect on SCP activity . The substrate specificity outlined in this work further supports the similarity between SCP and the cysteine proteases of the papain family . From the data regarding the identified or proposed natural substrates for SCP, it appears that this substrate specificity profile may also apply to the processing of mammalian and streptococcal protein targets by SCP. Infect Immun, 2001 Oct, 69(10), 6520 - 2 Regulation of Streptococcus gordonii sspB by the sspA gene product; El-Sabaeny A et al.; Streptococcus gordonii expresses two related adhesins, SspA and SspB, the genes for which are adjacent on the chromosome and are regulated independently . Although the adhesins are functionally similar, the sspA promoter is more active than that of sspB . In this study we show an additional role for SspA in the control of sspB activity . Gel shift and DNA footprinting assays demonstrate that the SspA protein binds to the sspB promoter and protects a region 233 to 264 bp upstream of the predicted -35 promoter element . The responsiveness of the sspB promoter to SspA was investigated with a promoter-cat reporter . Expression of the sspB promoter was reduced by over 60% in an SspA-deficient mutant of S . gordonii . These results indicate that expression of S . gordonii sspB is positively regulated by the sspA gene product. Infect Immun, 2001 Oct, 69(10), 6186 - 92 Proteins PblA and PblB of Streptococcus mitis, which promote binding to human platelets, are encoded within a lysogenic bacteriophage; Bensing BA et al.; The binding of platelets by bacteria is a proposed central mechanism in the pathogenesis of infective endocarditis . Platelet binding by Streptococcus mitis strain SF100 (an endocarditis isolate) was recently shown to be mediated in part by the surface proteins PblA and PblB . The genes encoding PblA and PblB are clustered with genes nearly identical to those of streptococcal phages r1t, 01205, and Dp-1, suggesting that pblA and pblB might reside within a prophage . To address this possibility, cultures of SF100 were exposed to either mitomycin C or UV light, both of which are known to induce the lytic cycle of many temperate phages . Both treatments caused a significant increase in the transcription of pblA . Treatment with mitomycin C or UV light also caused a substantial increase in the expression of PblA and PblB, as detected by Western blot analysis of proteins in the SF100 cell wall . By electron microscopy, phage particles were readily visible in the supernatants from induced cultures of SF100 . The phage, designated SM1, had a double-stranded DNA genome of approximately 35 kb . Southern blot analysis of phage DNA indicated that pblA and pblB were contained within the SM1 genome . Furthermore, Western blot analysis of phage proteins revealed that both PblA and PblB were present in the phage particles . These findings indicate that PblA and PblB are encoded by a lysogenic bacteriophage, which could facilitate the dissemination of these potential virulence determinants to other bacterial pathogens. Infect Immun, 2001 Oct, 69(10), 6102 - 9 Effect on polymorphonuclear cell function of a human-specific cytotoxin, intermedilysin, expressed by Streptococcus intermedius; Macey MG et al.; Streptococcus intermedius is a member of the normal flora of the mouth but is also an opportunistic pathogen associated with purulent infections at oral and nonoral sites . Intermedilysin (ILY) has been shown to be a cytolysin capable of generating pores in the cell membrane of erythrocytes demonstrable by electron microscopy . This effect has been shown to be specific for human cells . Since polymorphonuclear cells (PMNs) are the main cell involved in innate immunity we investigated the effect of purified intermedilysin from Streptococcus intermedius on PMN function . Active ILY at a concentration of 40 ng/microl caused a significant decrease in the number of intact PMNs after 60 min . The active cytolysin, when compared with heat-inactivated ILY, did not appear to be chemotactic for the PMNs but did cause an increase in intracellular calcium, with increased cell surface CD11b expression, metabolic burst, and phagocytosis of Staphylococcus aureus . These findings may have implications for the role of ILY in deep-seated abscesses. Infect Immun, 2001 Oct, 69(10), 6055 - 63 Population dynamics of Streptococcus mitis in its natural habitat; Hohwy J et al.; The purpose of this study was to examine the genetic structure of the typical commensal Streptococcus mitis biovar 1 in its natural habitat in the human oral cavity and pharynx and to investigate the role that selected microbial properties and host, spatial, and temporal factors play in determining the structure of the bacterial population . Consecutive samples were collected from buccal and pharyngeal mucosal surfaces of two infants, their four parents, and two elderly individuals over a period of approximately 1 year . A total of 751 isolates identified as S . mitis biovar 1 were typed by restriction endonuclease analysis (REA) and representative clones were typed by multilocus enzyme electrophoresis (MLEE) . The genetic diversity of the S . mitis biovar 1 isolates collected from single infant hosts over a period of 9 to 10 months was found to be between 0.69 and 0.76, which is considerably higher than that previously observed for intestinal populations of Escherichia coli . The study provides evidence of the existence of both transient and persistent clones in adult individuals . In the two infants, however, none of 42 demonstrated clones were detected on more than a single occasion . Statistical calculations showed that the ability to persist was not distributed at random in the S . mitis biovar 1 population . However, neither immunoglobulin A1 protease activity nor the ability to bind alpha-amylase from saliva was a preferential characteristic of persistent genotypes . In contrast to current concepts of climax ecosystems, the species niche in the habitat appears to be maintained predominantly by a succession of clones rather than by stable strains . Several lines of evidence suggest that the major origin of "new" clones is the many other habitats in the respiratory tract that are occupied by this species. Infect Immun, 2001 Oct, 69(10), 6030 - 7 Surface-expressed mig protein protects Streptococcus dysgalactiae against phagocytosis by bovine neutrophils; Song XM et al.; The mig gene of Streptococcus dysgalactiae, a major bovine mastitis pathogen, encodes two plasma protein-binding receptors, alpha2-macroglobulin (alpha2-M) and immunoglobulin G (IgG) . In this study, the mig gene from one S . dysgalactiae isolate was cloned and expressed in Escherichia coli . The IgG receptor region encoded by mig was conserved in 16 S . dysgalactiae strains . An isogenic mig mutant was constructed by allele replacement mutagenesis of the wild-type gene in S . dysgalactiae . The IgG-binding activity was lost in the mig mutant strain, whereas the alpha2-M receptor activity was still expressed but was detected only in the culture supernatant . In flow cytometry phagocytosis and bacterial-colony-counting bactericidal assays, the wild-type strain was found to be significantly more resistant to phagocytosis and killing by bovine neutrophils (PMNs) than the mig mutant strain when bacteria were preincubated with bovine serum . We therefore speculate that the Mig protein of S . dysgalactiae plays a role in virulence of the bacteria by binding to the plasma protein alpha2-M or IgG and thus preventing phagocytosis by bovine PMNs. Infect Immun, 2001 Oct, 69(10), 5997 - 6003 Protection against Streptococcus pneumoniae elicited by immunization with pneumolysin and CbpA; Ogunniyi AD et al.; The need for the development of cheap and effective vaccines against pneumococcal disease has necessitated the evaluation of common virulence-associated proteins of Streptococcus pneumoniae as potential vaccine antigens . In this study, we examined the capacity of active immunization with a genetic toxoid derivative of pneumolysin (PdB) and/or a fragment of choline binding protein A (CbpA; also known as PspC, Hic, and SpsA) to protect mice from intraperitoneal challenge with medium to very high doses of a highly virulent capsular type 2 pneumococcal strain, D39 . The median survival times for mice immunized with the individual protein antigens in different adjuvant combinations were significantly longer than those for mice that received the respective adjuvants alone . Mice immunized with CbpA alone were significantly better protected than mice immunized with PdB alone . Correspondingly, the median survival times for mice that were immunized with a combination of PdB and CbpA were significantly longer than those for mice that received PdB alone but not significantly different from those that received CbpA alone . Mice immunized with the protein antigens in a mixture of monophospholipid A (MPL) and aluminium phosphate (AlPO4) adjuvants had higher antibody titers than mice that received the antigens in AlPO4 alone . Mice immunized with PdB in MPL plus AlPO4 were also significantly better protected than mice that received PdB in AlPO4 alone. Acta Med Port, 2001 May-Jun, 14(3), 367 - 70 {Necrotizing fasciitis after varicella}; Goncalves E et al.; Necrotizing fasciitis is a rare and severe infection characterised by extremely rapid progressive involvement of the superficial fascias and deep dermal layers of the skin, with resultant vasculitis and necrosis . The authors present three clinical cases of necrotizing fasciitis; all three patients previously had varicella rash, rapid progressive spreading erythema with severe pain and toxic shock syndrome . Two patients had positive cultures of b-haemolytic streptococcus . Early stage differential diagnosis with celulitis, aggressive antibiotic treatment and pediatric intensive care support are essential . However, the main therapy is early extensive surgical approach involving all indurate areas, down to and including the muscle fascia. Oper Dent, 2001 Sep-Oct, 26(5), 445 - 50 Effect of adhesives on the inhibition of secondary caries around compomer restorations; Itota T et al.; This study evaluated the effect of adhesives on the inhibition of secondary caries around compomer restorations in vitro . Two adhesive systems with a Bis-GMA resin, Scotch bond Multi-purpose (MP) and Single Bond (SB), and one adhesive system with no Bis-GMA resin, F2000 compomer primer/adhesive (PA), were used prior to placement of the compomer (F2000), and non-fluoride releasing resin composite (Z100) was used as a control . Class V cavities prepared on extracted human premolars were restored with various combinations of materials: F2000/MP, F2000/SB, F2000/PA, Z100/MP, Z100/SB and Z100/PA . The restored teeth were incubated in bacterial medium containing sucrose with Streptococcus mutans for two weeks after storage for 14 days . On microradiographs, the radio-opaque layers adjacent to the F2000 restorations were thick and clear, while the layers in the Z100 restorations were unclear . In the F2000 restorations, the mean thickness of the radio-opaque layers in the PA group was significantly greater than that of the MP and SB groups . In fluoride-releasing measurement, F2000 coated with PA showed a significantly higher amount of fluoride release than MP and SB, and no significant difference in the amount of fluoride release from uncoated F2000 . These results indicated that applying an adhesive without Bis-GMA resin to compomer restoration has no suppressive effect on the fluoride release from compomer and might be beneficial for inhibiting secondary caries in vitro. Microbiology, 1995 Oct, 141(Pt 10), 2729 - 38 Polypeptides associated with tufts of cell-surface fibrils in an oral Streptococcus; Jameson MW et al.; Cells of the oral bacterium Streptococcus oralis CN3410 produce lateral tufts of cell-surface fibrils of two lengths . Treatment of cells with trypsin resulted in loss of the tufts and release of longer fibrils intact . SDS-PAGE analysis of trypsin extracts containing fibrils revealed two groups of high molecular mass polypeptides which were denoted group A (molecular mass 227-246 kDa) and group B (molecular mass 175-208 kDa) . Antibodies were raised to these two groups of trypsin-extracted polypeptides (TEPs) and to purified fibrils, and the reactivities of the three different antisera were found to be similar both on nitrocellulose blots of cell-surface polypeptides and in ELISA with whole cells . Similar patterns of TEPs were obtained from cells of a spontaneously derived mutant strain, KP34V, which lacked the short fibril components of tufts . Cells of strain KP34V had similar cell-surface hydrophobicity to strain CN3410 cells, and adhered to the same extent to parotid salivary pellicle or human buccal epithelial cells (BECs) as the wild-type cells . Trypsin treatment of strain CN3410 cells abolished their surface hydrophobicity and ability to adhere to BECs, but did not affect streptococcal cell binding to experimental salivary pellicle . Antibodies to TEPs or fibrils had no effect on cell adhesion to BECs or salivary pellicle . The results imply that the short fibril components of tufts are not involved in the cell adhesion properties tested . It is suggested that the TEPs are components of long fibrils, but they are not determinants of streptococcal cell adhesion to pellicle or to epithelial cells. Clin Infect Dis, 2001 Oct 15, 33(8), e93 - 6 Epub 2001 Sep 05. Emergence of rifampin-resistant Streptococcus pneumoniae as a result of antimicrobial therapy for penicillin-resistant strains; van Tilburg PM et al.; A multidrug-resistant strain of Streptococcus pneumoniae was isolated in The Netherlands during a nosocomial outbreak among 36 patients who mainly had chronic obstructive pulmonary disease . After the commencement of barrier nursing and short-term ceftriaxone-rifampin eradication therapy, the epidemic ceased . However, eradication therapy failed in 3 patients, and follow-up investigation of these patients showed the emergence of rifampin-resistant isolates. Pediatr Neurosurg, 2001 Aug, 35(2), 82 - 9 Intracranial complications of frontal sinusitis in children: Pott's puffy tumor revisited; Bambakidis NC et al.; The objective of the present study is to describe the diagnosis and treatment of intracranial complications of frontal sinusitis (Pott's puffy tumor) in a series of pediatric patients at our institution . A rare entity, Pott's puffy tumor has been reported in only 21 pediatric cases in the literature of the antibiotic era . The hospital records and radiographic files at Rainbow Babies and Childrens Hospital, Cleveland, Ohio, USA, over the previous 16 years were retrospectively reviewed in a search for patients with the diagnosis of Pott's puffy tumor, defined as scalp swelling and associated intracranial infection . There were 6 male patients and 1 female patient . Ages ranged from 11 to 18 years (median 14.5 years) . Intracranial infections consisted of epidural abscess in 5 patients, subdural empyema in 4 and brain abscess in 1 . Intraoperative cultures grew anaerobic organisms in 1 patient, microaerophilic streptococcus in 5 patients, Klebsiella species in 1 patient and Streptococcus pneumoniae in another . All patients presented with frontal scalp swelling, and other common symptoms included headache, fever, nasal drainage and frontal sinus tenderness . Five patients were treated with antibiotics prior to their presentation . Four patients presented with neurologic decompensation characterized by varying degrees of hemiparesis, obtundation, pupillary dilatation or aphasia . All patients underwent craniotomy and evacuation of the intracranial infection . Even severely impaired patients demonstrated full neurologic recovery . Despite the widespread use of antibiotics, neurosurgical complications of sinusitis continue to occur . A high degree of suspicion, along with prompt neurosurgical intervention and the use of appropriate antibiotics, can result in favorable outcomes in even the sickest patients . J Med Microbiol, 2001 Sep, 50(9), 828 - 32 Alterations to penicillin-binding proteins 1A, 2B and 2X amongst penicillin-resistant clinical isolates of Streptococcus pneumoniae serotype 23F from the nasopharyngeal flora of children; Ferroni A et al.; Various amino acid substitutions were identified in the three major penicillin-binding proteins (PBP1A, PBP2B and PBP2X) of eight clinical isolates of Streptococcus pneumoniae serotype 23F collected from children . The particular changes related to the level of penicillin resistance . Alterations were detected in an isolate with a penicillin MIC as low as 0.06 mg/L . These results confirm that the level of penicillin resistance in pneumococci reflects with sequential alterations of PBPs in clinical isolates. J Med Microbiol, 2001 Sep, 50(9), 812 - 21 Tumour necrosis factor-alpha causes an increase in blood-brain barrier permeability during sepsis; Tsao N et al.; Blood-brain barrier (BBB) permeability during sepsis with Escherichia coli or Streptococcus pneumoniae was examined in a mouse model and measured by a circulating beta-galactosidase tracer . The leakage of brain microvascular vessels during sepsis was confirmed by transmission electron microscopic examination of brain tissues stained with horseradish peroxidase . The increase of BBB permeability induced by E . coli and S . pneumoniae, which was maximal at 3 h and 12 h after injection, respectively, was transient because of rapid clearance of the bacteria from the blood . Tumour necrosis factor-alpha (TNF-alpha) was stained on microvascular vessels of the brain during sepsis and intravenous injection of recombinant TNF-alpha also increased the BBB permeability . The increase in BBB permeability induced by either E . coli or S . pneumoniae could be inhibited by anti-TNF-alpha antibody . It was concluded that circulating TNF-alpha generated during sepsis induced the increase in BBB permeability. Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 531 - 8 The impact of fermentative organisms on carbon flow in methanogenic systems under constant low-substrate conditions; Dollhopf SL et al.; We compared carbon flow under constant low-substrate conditions (below 20 microM glucose in situ) in laboratory-scale glucose-fed methanogenic bioreactors containing two very different microbial communities that removed chemical oxygen demand at similar rates . One community contained approximately equal proportions of spiral and cocci morphologies, while the other community was dominated by cocci . In the former bioreactor, over 50% of the cloned SSU rRNA genes and the most common SSU rDNA terminal restriction fragment corresponded to Spirochaetaceae-related sequences, while in the latter bioreactor over 50% of the cloned SSU rRNA genes and the most common SSU rDNA terminal restriction fragment corresponded to Streptococcus-related sequences . Carbon flow was assessed by measuring 14C-labeled metabolites derived from a feeding of {U-14C}glucose that did not alter the concentration of glucose in the bioreactors . Acetate and ethanol were detected in the Spirochaetaceae-dominated reactor, whereas acetate and propionate were detected in the Streptococcus-dominated reactor . A spirochete isolated from a Spirochaetaceae-dominated reactor fermented glucose to acetate, ethanol, and small amounts of lactate . Maximum substrate utilization assays carried out on fluid from the same reactor indicated that acetate and ethanol were rapidly utilized by this community . These data indicate that an acetate- and ethanol-based food chain was present in the Spirochaetaceae-dominated bioreactor, while the typical acetate- and propionate-based food chain was prevalent in the Streptococcus-dominated bioreactor. Salud Publica Mex, 2001 Jul-Aug, 43(4), 352 - 67 Meeting the challenge: prevention of pneumococcal disease with conjugate vaccines; Echaniz-Aviles IG et al.; Streptococcus pneumoniae is one of the leading causes of both invasive and noninvasive diseases in the pediatric population and continues to represent a significant public health burden worldwide . The increasing incidence of antibioticresistant strains of the pathogen has complicated treatment and management of the various pneumococcal disease manifestations . Thus, the best management strategy may be the prevention of pneumococcal diseases through vaccination . Although several pneumococcal conjugate vaccines have been clinically studied in infants and children, only a 7-valent conjugate vaccine (PNCRM7; Prevnar/Prevenar) is currently approved for the prevention of invasive disease . Vaccination with PNCRM7 is safe and effective in infants and young children . Routine vaccination with the conjugate vaccine could improve outcomes by safeguarding against the development of antibiotic-resistant strains of S . pneumoniae, thus simplifying the management of pneumococcal disease . Additionally, the overall costs associated with the treatment of pneumococcal diseases could be substantially reduced, particularly in developing countries . The time has come for fully applying this new advancement against S . pneumoniae, to benefit the children of the world . The English version of this paper is available at: http://www.insp.mx/salud/index.html Harefuah, 2001 Aug, 140(8), 704 - 5, 807 {Primary peritonitis due to Streptococcus pyogenes}; Shukha A et al.; Most infections causing by Streptococcus pyogenes are known to be putrid both in adults and children, for example laryngitis, impetigo, cellulitis, scarlet fever, septic shock, toxic shock syndrome etc . Their clinical importance relates to a number of causes: 1) about 20% of the population are S . pyogenes carriers; 2) the clinical course of infections is often severe or even fatal; 3) there is real danger of nosocomial outbursts; 4) a world-wide increase in both the rate and invasiveness of the microbe . Acute primary peritonitis due to S . pyogenes seems to be a rare clinical event . In order to stimulate physicians' attention to the issue, we decided to describe the case of a child with the primary peritonitis due to S . pyogenes . Early diagnosis and surgical intervention combined with the proper antibiotic treatment appeared to be life-saving. J Biol Chem, 2001 Nov 9, 276(45), 41790 - 6 Epub 2001 Sep 06. Similarities between complement-mediated and streptolysin S-mediated hemolysis; Carr A et al.; The oxygen-stable hemolysin streptolysin S (SLS) of Streptococcus pyogenes is encoded in part by the pel/sagA gene product . Antibodies to a synthetic peptide from the C terminus of the Pel/SagA open reading frame inhibited hemolysis mediated by both culture supernatants from multiple M serotypes of S . pyogenes isolates or a commercially available SLS preparation . Analysis of the SLS-mediated hemolytic reaction demonstrated that it was temperature- and concentration-dependent . Like complement-mediated hemolysis it conforms to the prediction of a one-hit mechanism of hemolysis . A number of intermediates in the SLS-mediated hemolysis of sheep erythrocytes could be distinguished . SLS could bind to erythrocytes below 17 degrees C; however, lysis could only occur at temperatures >23 degrees C . Following binding of SLS and washing, a papain-sensitive intermediate could be distinguished prior to insertion of the SLS complex into the erythrocyte membrane, which resulted in formation of a transmembrane pore and led to irreversible osmotic lysis of the cell . These intermediates were similar to those described previously during complement-mediated hemolysis. J Infect, 2001 May, 42(4), 272 - 7 Necrotizing fasciitis due to penicillin-resistant Streptococcus pneumoniae: case report and review of the literature; Ballon-Landa GR et al.; Necrotizing fasciitis (NF) is a life-threatening infection involving rapid necrosis of subcutaneous and fascial tissues . Streptococcus pneumoniae (SPN) soft tissue infection is exceedingly uncommon, reported primarily in patients with immunosuppression or other underlying conditions . We report a case of NF and septic shock in a healthy 32-year-old man, whose only predisposing factor was antecedent blunt trauma . Pathological examination and culture of the extensive tissue debridement were positive only for SPN . The serotype 9V isolate was penicillin (PCN)-resistant (MIC=2.0), and closely-related by pulse field gel electrophoresis and multilocus fingerprinting to clone France 9V-3, an important genetic reservoir for increasing PCN-resistance worldwide . This unique case has implications for our pathogenic under-standing and empiric management of NF . J Infect, 2001 May, 42(4), 235 - 42 Contribution of alveolar phagocytes to antibiotic efficacy in an experimental lung infection with Streptococcus pneumoniae; Calame W et al.; OBJECTIVES: The effect of cyclophosphamide-induced leukocytopenia on the cellular defence and on the efficacy of penicillin treatment in a Streptococcus pneumoniae pneumonia model in mice was studied . METHODS: The number of alveolar phagocytes was determined in broncho-alveolar lavage (BAL) fluid as well as the number of bacteria in both BAL fluid and homogenized lung tissue . RESULTS: Eighteen and 21 h after infection, leukocytopenic animals had significantly lower numbers of alveolar phagocytes than controls, while the numbers of bacteria in both BAL fluid and lungs were significantly higher . The number of bacteria was inversely related to the dose of penicillin and the number of alveolar macrophages . The number of alveolar granulocytes was inversely related to the dose of penicillin . CONCLUSIONS: Leukocytopenia due to cyclophosphamide impairs the cellular defence in the lung against Streptococcus pneumoniae and the dose of penicillin must be increased to compensate for the higher outgrowth of bacteria in these leukocytopenic mice, compared to normal animals . J Infect, 2001 Apr, 42(3), 210 - 2 Streptococcus pneumoniae: a rare skin pathogen? Kalima P, Riordan T. Streptococcus pneumoniae is a rare cause of skin infections in adults . We present three cases and a review of the literature on this infection . Nine of the 42 (21%) cases occurred in previously healthy individuals without predisposing conditions . The majority of cases (88%) had bacteraemia . More than half the cases (22/42, 52%) required surgical intervention in addition to antimicrobial therapy . Infection, 2001 Aug, 29(4), 234 - 6 Spinal epidural abscess due to Streptococcus pneumoniae in an HIV-infected adult; Younus F et al.; Spinal epidural abscess (SEA) due to Streptococcus pneumoniae is rare and has never been reported in an HIV-infected patient, despite the higher risk of invasive disease in this group . We describe here the first case of pneumococcal epidural abscess, presenting with fever and back pain in a 60-year-old man infected with HIV . Blood cultures were positive for S . pneumoniae and magnetic resonance imaging (MRI) confirmed the suspicion of diskitis and SEA at the L4-S1 level . The patient was successfully treated with iv ceftriaxone without surgical intervention . The clinical characteristics of this case are compared with existing literature on pneumococcal SEA. Infection, 2001 Aug, 29(4), 201 - 4 Efficacy of clarithromycin in preventing viridans streptococcal bacteremia following autologous stem cell transplantation; Naqvi B et al.; BACKGROUND: In a study involving 200 patients, we previously found that 17.5% of patients developed viridans streptococcal (VS) bacteremia following autologous peripheral blood stem cell transplantation (aPBSCT) when ciprofloxacin or ciprofloxacin plus ampicillin was used for prophylaxis . PATIENTS AND METHODS: A retrospective evaluation of 100 consecutive recipients of aPBSCT was conducted to ascertain the incidence and outcome of VS bacteremia when a combination of ciprofLoxacin and clarithromycin was utilized for antimicrobiaL prophylaxis following transplantation . The 200 patients from our previous study, in which ciprofloxacin alone or ciprofloxacin with ampicillin was used for prophylaxis, were combined with the current group for the purpose of statistical analysis . RESULTS: Streptococcus mitis was isolated from the blood of five individuals at a median of 5 days following stem cell infusion . Each of these patients was neutropenic and presented with fever . Three isolates demonstrated intermediate resistance to macrolides in vitro . However, all episodes of bacteremia were treated successfully with systemic antibiotic therapy . CONCLUSION: Age, duration of neutropenia, type of underlying malignancy and type of conditioning chemotherapy regimen failed to have a significant impact on subsequent VS bacteremia . Only female sex and use of ciprofloxacin without clarithromycin as antimicrobiaL prophyLaxis predicted a significantly increased risk of VS bacteremia in both univariate and Logistic regression analyses. Clin Experiment Ophthalmol, 2001 Aug, 29(4), 260 - 1 Fulminant orbital cellulitis with complete loss of vision; Connell B et al.; Bacterial orbital cellulitis is a condition that rarely presents with complete loss of vision . A case is reported of a 69-year-old man who presented with fulminant onset of proptosis, significant ophthalmoplegia and no perception of light . Computed tomography showed no evidence of paranasal sinus disease . Despite treatment with intravenous flucloxacillin, ceftriaxone and metronidazole, and later, penicillin after Streptococcus pyogenes was grown from tissue culture, there was no improvement in vision; however ocular motility returned to normal. Odontostomatol Trop, 2001 Jun, 24(94), 19 - 22 {Septicemia of dental origin and post-extraction coma . Apropos of 3 cases}; Ngapeth-Etoundi M et al.; The authors report 3 cases of septicaemia and coma post dental extraction . These 3 patients had dental extraction in the same confessional dental clinic . After these acts the situation were so severe and they came to central hospital in Yaounde . Infections from dental origin proven by blood culture necessitate an early antibiotherapy and treatment, which propriate and sustain, but adapted to the antibiogram . The streptococcus of group F and Klebsiella pneumonia were challenged . One of the cases was negative . The presence of common signs (fever, tachycardia, chill) need the search for a secondary localisation which can be determine by X-ray. J Allergy Clin Immunol, 2001 Sep, 108(3), 424 - 9 Allergic inflammation enhances bacterial sinusitis in mice; Blair C et al.; BACKGROUND: Although it is not proven, one factor considered important in the development of sinusitis is allergic rhinitis . OBJECTIVE: The purpose of this study was to determine whether ongoing allergic rhinitis enhances the infection and inflammation associated with Streptococcus pneumoniae acute sinus infection . METHODS: BALB/c mice were sensitized to ovalbumin by intraperitoneal injection . After infection of the sinuses by S pneumoniae, either with or without concomitant administration of ovalbumin to induce allergic inflammation, mice were killed at various times and their heads were prepared for histologic evaluation of the sinuses . RESULTS: Mice became allergic to ovalbumin and developed eosinophilia in the sinus and lung cavities in response to ovalbumin administration to each of the respective cavities . In comparison with controls, the mice with ongoing nasal allergic inflammation that were inoculated with S pneumoniae had significantly more bacteria recovered at sacrifice and had significantly more inflammation, as indicated by neutrophil, eosinophil, and mononuclear influx into the sinus mucosa . The percentage of the sinus area occupied by neutrophil clusters was also increased after infection in the allergic mice in comparison with the control mice . CONCLUSION: Our data demonstrate that mice can be sensitized to ovalbumin and develop a localized allergic reaction in the skin, nose, or lung . An ongoing local allergic response augments bacterial infection in these animals . We also demonstrate that allergic sensitization alone, allergen exposure alone, or an allergic response at a distal site, the lung, does not augment the sinus infection. J Bacteriol, 2001 Oct, 183(19), 5709 - 17 Genome of the bacterium Streptococcus pneumoniae strain R6; Hoskins J et al.; Streptococcus pneumoniae is among the most significant causes of bacterial disease in humans . Here we report the 2,038,615-bp genomic sequence of the gram-positive bacterium S . pneumoniae R6 . Because the R6 strain is avirulent and, more importantly, because it is readily transformed with DNA from homologous species and many heterologous species, it is the principal platform for investigation of the biology of this important pathogen . It is also used as a primary vehicle for genomics-based development of antibiotics for gram-positive bacteria . In our analysis of the genome, we identified a large number of new uncharacterized genes predicted to encode proteins that either reside on the surface of the cell or are secreted . Among those proteins there may be new targets for vaccine and antibiotic development. FEMS Microbiol Ecol, 1996 Jul, 20(3), 155 - 62 Survival of potentially pathogenic human-associated bacteria in the rhizosphere of hydroponically grown wheat; Morales A et al.; Plants may serve as reservoirs for human-associated bacteria (H-AB) in long-term space missions containing bioregenerative life support systems . The current study examined the abilities of five human-associated potential pathogens, Pseudomonas aeruginosa, Pseudomonas cepacia, Staphylococcus aureus, Streptococcus pyogenes and Escherichia coli, to colonize and grow in the rhizosphere of hydroponically grown wheat, a candidate crop for life support . All of these bacteria have been recovered from past NASA missions and present potential problems for future missions . The abilities of these organisms to adhere to the roots of axenic five-day-old wheat (Triticum aestivum L . cv . Yecora rojo) were evaluated by enumeration of the attached organisms after a one hour incubation of roots in a suspension (approximately 10(8) cfu ml-1) of the H-AB . Results showed that a greater percentage of P . aeruginosa cells adhered to the wheat roots than the other four H-AB . Similarly incubated seedlings were also grown under attempted axenic conditions for seven days to examine the potential of each organism to proliferate in the rhizosphere (root colonization capacity) . P . cepacia and P . aerogiunosa showed considerable growth, E . coli and S . aureus showed no significant growth, and S . pyogenes died off in the wheat rhizosphere . Studies examining the effects of competition on the survival of these microorganisms indicated that P . aeruginosa was the only organism that survived in the rhizosphere of hydroponically grown wheat in the presence of different levels of microbial competition. Microbiology, 2001 Sep, 147(Pt 9), 2469 - 77 H(2)O(2)-nonproducing Streptococcus pyogenes strains: survival in stationary phase and virulence in chronic granulomatous disease; Saito M et al.; The production of hydrogen peroxide (H(2)O(2)) and related phenotypes were studied with Streptococcus pyogenes strains isolated from cases of pharyngitis or severe group A streptococcal infections . Of the 46 strains examined (34 from severe infections and 12 from pharyngitis cases), 25 strains accumulated H(2)O(2) in the culture medium when grown under glucose-limited, aerobic conditions, whereas the rest of the strains did not . There was no correlation between these traits and the type of disease from which each strain had been isolated . The H(2)O(2)-nonproducing strains tested in this study belonged to T type 3 or T type 12 . The accumulation of H(2)O(2) started when the culture reached the late exponential phase . A rapid loss of cell viability accompanied H(2)O(2) accumulation but was completely prevented by the addition of a catalase, indicating that the lethality was actually caused by H(2)O(2) . Cells of H(2)O(2)-nonproducing strains were resistant to killing by phagocytes from patients with chronic granulomatous disease (CGD), whereas those of H(2)O(2)-producing strains were subject to killing . Subcutaneous inoculation of 10(5) c.f.u . H(2)O(2)-nonproducing S . pyogenes strains into the hind footpads of CGD mice provoked more prominent swelling of the footpad than did H(2)O(2)-producing strains . The mortality rate in the CGD mice infected with the H(2)O(2)-nonproducing strains was higher than that produced by the H(2)O(2)-producing strains . It is suggested that H(2)O(2)-nonproducing S . pyogenes strains are prevalent in humans and that they may be a potential threat to the health of CGD patients. Vaccine, 2001 Sep 14, 19(32), 4780 - 90 Efficacy of pneumococcal polysaccharide vaccine in immunocompetent adults: a meta-analysis of randomized trials; Cornu C et al.; The use of pneumococcal polysaccharide vaccine (PPV) is low in some countries, maybe because of doubts regarding its efficacy . This meta-analysis aims at combining evidence from randomized trials of PPV assessing its efficacy in preventing Streptococcus pneumoniae related diseases in immunocompetent adults . In the fourteen trials totalling 48,837 patients retrieved, PPV prevents definite pneumococcal pneumonia by 71%, presumptive pneumococcal pneumonia by 40%, and mortality due to pneumonia by 32%, but not all-cause pneumonia or death . No preventive effect was seen in the subgroup of patients aged 55 years or more, possibly due to a lack of statistical power. Prev Vet Med, 2001 Oct 11, 51(3-4), 307 - 16 Application of the California mastitis test in intramammary Streptococcus agalactiae and Staphylococcus aureus infections of camels (Camelus dromedarius) in Kenya; Younan M et al.; A study was conducted on 207 lactating camels in six herds in Kenya to evaluate the California mastitis test (CMT) for the detection of intramammary infections (IMIs) caused by Streptococcus agalactiae and Staphylococcus aureus and to investigate the prevalence of both the pathogens in the camel udder . IMI with S . agalactiae was found in 12% of all camels sampled . IMI with S . aureus was present in 11% of all camels sampled . The herd-level prevalence of IMI varied between 0 and 50% for S . agalactiae and between 0 and 13% for S . aureus . Longitudinal observations over 10-12 months confirmed persistent infections for both pathogens . Observations in one herd suggested that camel pox was a contributing factor in spreading and exacerbating S . agalactiae udder infections.The CMT had quarter-level sensitivities of 77 and 68% for S . agalactiae and S . aureus in camels, respectively . The CMT specificities were 91% for both the pathogens. Respir Physiol, 2001 Oct, 128(1), 23 - 31 The role of immunity in susceptibility to respiratory infection in the aging lung; Meyer KC; Respiratory tract infections, particularly pneumonia, are a leading cause of death in persons 65 years or older in both developed and developing countries . Because many attributes of immunity wane with advancing age, the elderly may be more susceptible to respiratory infections, even if they appear to be in good health . A decline in the ability of lymphoid tissues to mount an antigen-specific response (adaptive immunity) to specific microorganisms such as influenza virus or Streptococcus pneumoniae is thought to be an important factor in increasing susceptibility to respiratory tract infection with advancing age . However, abnormalities in innate immunity may also contribute to increased susceptibility to respiratory infections and have been poorly characterized in the elderly . Although changes in immune parameters such as T cell subsets and immunoglobulin concentrations have been observed in respiratory secretions from older healthy individuals compared to younger subjects, the significance of these changes for protective immunity in the lung is unknown . The incidence of pneumonia may be lessened by measures such as optimizing treatment of comorbid conditions, optimizing nutrition, and addressing swallowing disorders . The use of vaccines directed against the influenza virus and S . pneumoniae appears to have made an impact on the degree of morbidity and mortality, and perhaps, the incidence, of community-acquired pneumonia . However, better stimulation of specific immune responses with improved vaccines and more widespread use of these vaccines for protection of elderly individuals are needed. Biochem J, 2001 Sep 15, 358(Pt 3), 529 - 38 Carbohydrate specificity of a galectin from chicken liver (CG-16); Wu AM et al.; Owing to the expression of more than one type of galectin in animal tissues, the delineation of the functions of individual members of this lectin family requires the precise definition of their carbohydrate specificities . Thus, the binding properties of chicken liver galectin (CG-16) to glycoproteins (gps) and Streptococcus pneumoniae type 14 polysaccharide were studied by the biotin/avidin-mediated microtitre-plate lectin-binding assay and by the inhibition of lectin-glycan interactions with sugar ligands . Among 33 glycans tested for lectin binding, CG-16 reacted best with human blood group ABO (H) precursor gps and their equivalent gps, which contain a high density of D-galactopyranose(beta1-4)2-acetamido-2-deoxy-D-glucopyranose {Gal(beta1-4)GlcNAc} and Gal(beta1-3)GlcNAc residues at the non-reducing end, but this lectin reacted weakly or not at all with A-,H-type and sialylated gps . Among the oligosaccharides tested by the inhibition assay, the tri-antennary Gal(beta1-4)GlcNAc (Tri-II) was the best . It was 2.1x10(3) nM and 3.0 times more potent than Gal and Gal(beta1-4)GlcNAc (II)/Gal(beta1-3)GlcNAc(beta1-3)Gal(beta1-4)Glc (lacto-N-tetraose) respectively . CG-16 has a preference for the beta-anomer of Gal at the non-reducing end of oligosaccharides with a Gal(beta1-4) linkage >Gal(beta1-3)> or =Gal(beta1-6) . From the results, it can be concluded that the combining site of this agglutinin should be a cavity type, and that a hydrophobic interaction in the vicinity of the binding site for sugar accommodation increases the affinity . The binding site of CG-16 is as large as a tetrasaccharide of the beta-anomer of Gal, and is most complementary to lacto-N-tetraose and Gal(beta1-4)GlcNAc related sequences. An Med Interna, 2001 Jul, 18(7), 379 - 80 {Endophthalmitis and multiple brain abscess in a patient with endocarditis due to Streptococcus agalactiae}; Pena Jimenez D et al.; Endogenous endophthalmitis is a rare disease caused by hematogenic germ spread from an internal focus . Infections due to Streptococcus agalactiae are infrequent in adults although new cases had been described recently associated to inmunodepression . We present a patient with endocarditis due to Streptococcus agalactiae, endophthalmitis and multiple brain abscess . We also review the literature. Pediatrics, 2001 Sep, 108(3), 575 - 83 A community intervention trial to promote judicious antibiotic use and reduce penicillin-resistant Streptococcus pneumoniae carriage in children; Belongia EA et al.; OBJECTIVE: Inappropriate use of antibiotics is common in primary care, and effective interventions are needed to promote judicious antibiotic use and reduce antibiotic resistance . The objective of this study was to assess the impact of parent and clinician education on pediatric antibiotic prescribing and carriage of penicillin-nonsusceptible Streptococcus pneumoniae in child care facilities . METHODS: A nonrandomized, controlled, community intervention trial was conducted in northern Wisconsin Clinicians . Clinic staff received educational materials and small-group presentations; materials were distributed to parents through clinics, child care facilities, and community organizations . Prescribing data were analyzed for 151 clinicians who provided primary pediatric care; nasopharyngeal carriage of penicillin-nonsusceptible S pneumoniae was assessed for 664 children in the baseline period (January-June 1997) and for 472 children in the postintervention period (January-June 1998) . RESULTS: The median number of solid antibiotic prescriptions per clinician declined 19% in the intervention region and 8% in the control region . The median number of liquid antibiotic prescriptions per clinician declined 11% in the intervention region, compared with an increase of 12% in the control region . Retail antibiotic sales declined in the intervention region but not in the control region . Among participating children in child care facilities, there were no significant differences in antibiotic use or penicillin-nonsusceptible S pneumoniae colonization between the intervention and control regions . CONCLUSIONS: A multifaceted educational program for clinicians and parents led to community-wide reductions in antibiotic prescribing, but in child care facilities, there was no apparent impact on judicious antibiotic use or colonization with drug-resistant S pneumoniae . Longer follow-up time or greater reductions in antibiotic use may be required to identify changes in the pneumococcal susceptibility. J Antimicrob Chemother, 2001 Sep, 48(3), 441 - 4 Antibiotic use and resistance of Streptococcus pneumoniae in The Netherlands during the period 1994-1999; de Neeling AJ et al.; Antibiotic use in The Netherlands during the period 1994-1999 is described in relation to the resistance of routine isolates of Streptococcus pneumoniae . The average antibiotic use in the study period was 3.4 defined daily doses per 1000 persons per day (DDD/1000/day) penicillins, 0.066 DDD/1000/day beta-lactams other than penicillins, 2.3 DDD/1000/day tetracyclines and 0.71 DDD/1000/day trimethoprim and sulphonamides, without apparent rise or decline . In contrast, the use of macrolides doubled from 0.51 DDD/1000/day in 1994 to 1.0 DDD/1000/day in 1997 and stayed at 1.07 DDD/1000/day in 1998 and 1999 . In 1994 the first pneumococci isolated from patients showed 0.7% resistance to penicillin (intermediate plus full resistance), 2.5% to erythromycin, 4.2% to co-trimoxazole and 4.7% to tetracycline . In 1999 first isolates showed 1.5% resistance to penicillin, 3.8% to erythromycin, 4.4% to co-trimoxazole and 6.6% to tetracycline . The modest but significant rise in the resistance to erythromycin may have been caused by the increased use of macrolides in the years 1994-1997 . The rise in resistance to penicillin seemed not to be related to increased beta-lactam use. J Antimicrob Chemother, 2001 Sep, 48(3), 411 - 6 In vitro selection of resistance to clindamycin related to alterations in the attenuator of the erm(TR) gene of Streptococcus pyogenes UCN1 inducibly resistant to erythromycin; Fines M et al.; A clinical isolate of Streptococcus pyogenes UCN1 intermediate to erythromycin (MIC 1 mg/L) and susceptible to clindamycin (MIC 0.03 mg/L) harboured an inducible erm(TR) gene encoding a ribosomal methylase . We have selected in vitro, in the presence of concentrations of clindamycin ranging from 0.12 to 1 mg/L, one-step mutants that are highly resistant to this antibiotic (MIC 64 mg/L) at a frequency of 10(-7) . By contrast, in an erythromycin-susceptible strain of S . pyogenes UCN5, mutants could be selected only by a low concentration of clindamycin (0.12 mg/L) at a frequency of 10(-9) . Clindamycin resistance in four of six S . pyogenes UCN1 mutants was associated with deletions of 163 and 6 bp, as well as a tandem duplication of 101 bp in the regulatory sequence of the erm(TR) gene . The role of these structural alterations in clindamycin resistance was demonstrated by cloning the erm(TR) gene from the wild-type and mutant strains in Escherichia coli DB10, a mutant susceptible to macrolides . Clindamycin resistance was expressed only when the erm(TR) gene was preceded by an altered attenuator . Mutations could lead to the formation of mRNA secondary structures accounting for the accessibility of the ribosome-binding site and the initiation codon of the ErmTR methylase to the ribosomes, and subsequently for the translation of the erm(TR) transcripts . The easy selection in one step of mutants resistant to high levels of clindamycin by concentrations of this antibiotic ranging from four to 40 times the MIC leads us to recommend caution in the use of clindamycin therapy in group A Streptococcus infections. J Antimicrob Chemother, 2001 Sep, 48(3), 407 - 9 In vitro activity of a novel ketolide ABT-773 against invasive strains of Streptococcus pneumoniae; Weiss K et al.; New ketolides such as ABT-773 are a promising group of antibiotics in an era of increasing antibiotic resistance . We tested 704 invasive strains of Streptococcus pneumoniae collected from 1990 to 1998 . Overall resistance was 8.3, 4.6, 4.5 and 3.6% for penicillin, cefuroxime, erythromycin and clarithromycin, respectively . By using a recommended breakpoint for susceptibility of <0.5 mg/L, no strains showed reduced susceptibility to ABT-773 . ABT-773 was very active against all penicillin-resistant strains (MIC > 2 mg/L, with a mean geometric mean <0.06 mg/L), and against all 33 erythromycin-resistant strains, irrespective of the mode of resistance {mef- or erm(B)-mediated} . ABT-773 is a very active and promising agent against invasive strains of S . pneumoniae, including multiresistant strains. J Antimicrob Chemother, 2001 Sep, 48(3), 365 - 74 Single- and multi-step resistance selection study of gemifloxacin compared with trovafloxacin, ciprofloxacin, gatifloxacin and moxifloxacin in Streptococcus pneumoniae; Nagai K et al.; The ability of sequential subcultures in subinhibitory concentrations of gemifloxacin, trovafloxacin, ciprofloxacin, gatifloxacin and moxifloxacin to select resistant mutants was studied in 16 pneumococci {eight with ciprofloxacin MICs (mg/L) 0.25-1; four with 8-16; four with 16-32} . Subculturing was done 50 times, or until mutants with elevated MICs (> or = 4 x) to the selecting drug emerged . Subculturing in gemifloxacin selected six resistant mutants (gemifloxacin MICs 2 mg/L); trovafloxacin selected nine (trovafloxacin MICs 2-4 mg/L); ciprofloxacin selected 11 (ciprofloxacin MICs 8-128 mg/L); gatifloxacin selected 13; and moxifloxacin selected 12 (gatifloxacin or moxifloxacin MICs 2-16 mg/L) . DNA sequencing showed that most mutants had mutations in ParC at Ser-79 or Asp-83 and in GyrA at Ser-81 or Glu-85; some mutants also had mutations in ParE or GyrB . Some new mutations were found in ParE or GyrB that have not yet been reported; GyrB mutation might be associated with moxifloxacin resistance . Both DNA gyrase and topoisomerase IV were thought to be the target of gemifloxacin; gemifloxacin also selected mutants with single modifications in gyrA, parC or parE alone among derived mutants by repeated exposure to subinhibitory concentrations of fluoroquinolones . In the presence of reserpine, most mutants had lower MICs of ciprofloxacin and gemifloxacin (4-32 x), and gatifloxacin (4-8 x), suggesting an efflux mechanism; none had lower trovafloxacin and moxifloxacin MICs . All quinolones tested selected for resistance; judicious use and proper dosing will be necessary to avoid resistance selection of newer broad-spectrum fluoroquinolones. Clin Microbiol Infect, 2001 Jul, 7(7), 362 - 6 In vitro activity of midecamycin diacetate, a 16-membered macrolide, against Streptococcus pyogenes isolated in France, 1995-1999; Schlegel L et al.; OBJECTIVE: To compare the in vitro activity of midecamycin diacetate to that of five other macrolides (erythromycin, clarithromycin, roxithromycin, azithromycin, and josamycin) and of clindamycin against 146 clinical isolates of Streptococcus pyogenes, with regard to three different phenotypes of erythromycin resistance . METHODS: Susceptibility pattern and resistance phenotype were determined by disk diffusion method and double disk test . Minimal inhibitory concentrations of antibiotics were obtained by the agar dilution method and evaluated according to the recommendations of the 'Comite de l'Antibiogramme de la Societe Francaise de Microbiologie' (CA-SFM) . The major determinants of erythromycin resistance in S . pyogenes (ermB, ermTR and mefA genes) were investigated by specific amplification protocols . RESULTS: Most of the isolates of S . pyogenes collected during 1995-99 were susceptible to midecamycin (93.8%), erythromycin (90.4%), clarithromycin (93.2%), roxithromycin (91.8%), azithromycin (88.4%), josamycin (94.5%), and clindamycin (94.5%) . According to the CA-SFM criteria, 132 of the 146 isolates studied were susceptible to erythromycin (MICs < or = 1 mg/L), four were intermediate (MICs 2-4 mg/L), and 10 were resistant (MICs > 4 mg/L) . Only nine isolates were midecamycin resistant (MICs > 4 mg/L), and the others were susceptible . The increased activity of midecamycin (MIC90 < or = 0.06 mg/L), as compared to erythromycin (MIC90 = 0.5 mg/L) and to other 14- or 15-membered macrolides, was related to the absence of the ermB determinant in seven isolates which displayed an efflux phenotype (five isolates) or an inducible resistance phenotype due to an ermTR determinant (two isolates) . CONCLUSION: Midecamycin diacetate is active against most S . pyogenes strains isolated in France and may represent an attractive alternative to the treatment of streptococcal infections due to resistant isolates with efflux of erythromycin. J Infect, 2001 Feb, 42(2), 116 - 9 Prevalence of liver disease in patients with Streptococcus bovis bacteraemia; Gonzlez-Quintela A et al.; BACKGROUND: The relationship between Streptococcus bovis bacteraemia and gastrointestinal disease (mainly colon cancer) is well known . Patients with advanced liver disease are prone to bacteraemia . Less attention has been paid to the association between liver disease and Streptococcus bovis bacteraemia in the literature . AIMS: To evaluate the prevalence of liver disease in patients with S . bovis bacteraemia . PATIENTS AND METHODS: Twenty-two episodes of S . bovis bacteraemia in 20 adults (13 males and seven females, with a median age of 61 years, range 32-94 years) were detected in a single hospital over a 7-year period . Ten of them had endocarditis . Patients' clinical records were reviewed, with special focus on underlying liver and gastrointestinal disease . RESULTS: Eleven patients (55%) had a chronic liver disease . Nine of them were cirrhotics . Ten patients had a history of chronic alcohol abuse, and four patients had hepatitis C virus antibodies (associated with alcohol abuse in three cases) . Large bowel disease was present in six out of 13 evaluable patients (adenocarcinoma in three cases) . Patients with liver disease were younger than patients without it . Mortality related to S . bovis bacteraemia was particularly high among patients with advanced liver disease (Child-Pugh state C) . Bacteraemia recurred two times in one alcoholic cirrhotic, who was diagnosed as having a Dukes-B colon cancer 4.5 years after the first episode of S . bovis bacteraemia . CONCLUSIONS: In our area, S . bovis bacteraemia is frequently associated with chronic liver disease . Liver disease may be a predisposing factor for S . bovis bacteraemia. Arch Otolaryngol Head Neck Surg, 2001 Aug, 127(8), 985 - 90 C57Bl/6 and BALB/c mice have similar neutrophil response to acute Streptococcus pneumoniae sinus infections; Gabr U et al.; BACKGROUND: Previous investigations have shown that mice with a tendency toward a T(H)1 or T(H)2 lymphocyte response manifest different reactions to inoculation with the parasite Leishmania major . BALB/c mice (with a tendency for a T(H)2 response) showed evidence of systemic infection, whereas C57Bl/6 mice (with a tendency for a T(H)1 response) showed only a local reaction . OBJECTIVE: To investigate whether BALB/c and C57Bl/6 mice respond differently to acute bacterial infection of the sinuses . METHODS: We inoculated the nasal cavities of C57Bl/6 and BALB/c mice with Streptococcus pneumoniae (type ATCC59), or with broth as a control . The mice were humanely killed 2, 5, 10, and 14 days after inoculation . Their heads were fixed, decalcified, and embedded in paraffin blocks . Sections were stained with hematoxylin and eosin, and the degree of inflammation was quantified by the number of neutrophils per square millimeter of the sinus mucosa and the percentage of the sinus cavity occupied by neutrophil clusters . RESULTS: Both groups of mice showed evidence of inflammation that was significantly greater than controls (P =.01), with no difference between groups . There was a correlation between the number of neutrophils per square millimeter in the sinus mucosa and the percentage of neutrophil clusters (C57Bl/6 mice, r = 0.37, P<.001; BALB/c mice, r = 0.20, P<.001) . In the infected mice, the number of infiltrating neutrophils was significantly greater (P<.001) in anatomically lower (dependent) areas of the sinuses compared with the upper areas . CONCLUSION: Unlike leishmaniasis, acute bacterial sinusitis is not affected by the tendency of the host to favor either a T(H)1 or T(H)2 response. AIDS Patient Care STDS, 2001 May, 15(5), 263 - 9 Successful prophylaxis against Pneumocystis carinii pneumonia in HIV-infected children using smaller than recommended dosages of trimethoprim-sulfamethoxazole; Fisher RG et al.; Prophylaxis against Pneumocystis carinii pneumonia (PCP) is an essential part of the management of children with human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS) . No dose-ranging studies were ever performed; therefore, the amount of trimethoprim-sulfamethoxazole (TMP-SMX) needed to suppress PCP in children with HIV/AIDS is not known . The dose recommended by the Centers for Disease Control (CDC) has been thought to be just above the threshold needed for prevention, based on anecdotal breakthrough PCP in cancer patients who were improperly dosed . We have been giving prophylaxis based on body weight rather than surface area, and this, combined with growth of our children, has led to a large experience with dosages lower than the currently recommended 150 mg/m2 . The medical records of children with HIV who met CDC guidelines for institution of PCP prophylaxis were reviewed . To ascertain the per square meter (m2) dosage each child was receiving, body surface area was calculated from height and weight measurements . Dosages were recalculated every 6 months and at each dosage change . Data regarding PCP infection, bacterial infections, and side effects of TMP-SMX were extracted . Data were compiled from 1,719.5 child-months of TMP-SMX prophylaxis, including 1,532.5 child-months below the currently recommended dose . Sixty-seven percent of our child-months were at or below two-thirds the CDC recommended dose . There were no cases of proven or suspected PCP . Incidence of other serious bacterial infections was low . Bacteremia and sepsis with Streptococcus pneumoniae was the most common proven bacterial infection, at a rate of 5.5 episodes per 100 child-years . The incidence of bacterial infection did not vary by the dose of TMP-SMX . TMP-SMX prophylaxis was well tolerated; most reactions were mild and self-limited and did not recur with re-institution of the drug . Only 6.1% of this cohort had TMP-SMX prophylaxis discontinued due to perceived toxicity . These data show that the currently recommended dose of TMP-SMX (150 mg/m2) may not be required to prevent PCP in children with HIV/AIDS . The drug is well tolerated at all dosage levels . The incidence of serious bacterial infection in this cohort of patients did not depend upon the amount of TMP-SMX prescribed . A prospective, controlled clinical trial of low-dose TMP-SMX for children with HIV infection is warranted. Eur Respir J, 2001 Aug, 18(2), 362 - 8 A prospective comparison of nursing home acquired pneumonia with community acquired pneumonia; Lim WS et al.; Nursing home acquired pneumonia (NHAP) is thought to be clinically distinct from community acquired pneumonia (CAP) . This observation, based on studies conducted mainly in North America, may not be relevant in countries with a different healthcare system . The authors describe an 18-month prospective cohort study of 437 patients admitted to hospital with CAP, 40 (9%) of whom came from nursing homes . Detailed microbiological tests were performed in a subset of patients over 12 months . Patients with NHAP were less likely to have a productive cough (odds ratio (OR) 0.4, p=0.02) or pleuritic pain (OR 0.1, p=0.03), but they were more likely to be confused (OR 2.6, p<0.001) . They had poorer functional status (p<0.001) and more severe disease (p=0.03) . Mortality was higher compared to CAP (53% versus 13%), but this was mainly explained by prior functional status (OR 0.5, after adjustment for functional status) . Pathogens were identified in 68% of 22 NHAP and 80% of 44 matched CAP patients . Streptococcus pneumoniae was the most common (55% NHAP, 43% CAP) . Atypical pathogens, enteric Gram negative bacilli and Staphylococcus aureus were uncommon . In conclusion, differences in functional status accounted for the increased mortality in nursing home acquired pneumonia compared to community acquired pneumonia . The pathogens implicated were similar . No grounds for a difference in choice of empirical antibiotics were apparent. J Biol Chem, 2001 Nov 2, 276(44), 41407 - 16 Epub 2001 Aug 29. Hyaluronan binding and degradation by Streptococcus agalactiae hyaluronate lyase; Li S et al.; Streptococcus agalactiae hyaluronate lyase is a virulence factor that helps this pathogen to break through the biophysical barrier of the host tissues by the enzymatic degradation of hyaluronan and certain chondroitin sulfates at beta-1,4 glycosidic linkages . Crystal structures of the native enzyme and the enzyme-product complex were determined at 2.1- and 2.2-A resolutions, respectively . An elongated cleft transversing the middle of the molecule has been identified as the substrate-binding place . Two product molecules of hyaluronan degradation were observed bound to the cleft . The enzyme catalytic site was identified to comprise three residues: His(479), Tyr(488), and Asn(429) . The highly positively charged cleft facilitates the binding of the negatively charged polymeric substrate chain . The matching between the aromatic patch of the enzyme and the hydrophobic patch of the substrate chain anchors the substrate chain into degradation position . A pair of proton exchanges between the enzyme and the substrate results in the cleavage of the beta-1,4 glycosidic linkage of the substrate chain and the unsaturation of the product . Phe(423) likely determines the size of the product at the product release side of the catalytic region . Hyaluronan chain is processively degraded from the reducing end toward the nonreducing end . The unsulfated or 6-sulfated regions of chondroitin sulfate can also be degraded in the same manner as hyaluronan. J Clin Microbiol, 2001 Sep, 39(9), 3316 - 20 Pneumococcal carriage in children in The Netherlands: a molecular epidemiological study; Bogaert D et al.; In 1999, Engelen and coworkers investigated colonization in Amsterdam among 259 children attending 16 day-care centers (DCCs) and among 276 children who did not attend day-care centers (NDCCs) . A 1.6- to 3.4-fold increased risk for nasopharyngeal colonization was observed in children attending DCCs compared with NDCC children, while no difference in antibiotic resistance was found between groups . The serotype and genotype distributions of 305 nasopharyngeal Streptococcus pneumoniae isolates of the latter study were investigated . The predominant serotypes in both the DCC and the NDCC groups included 19F (19 and 18%, respectively), 6B (14 and 16%, respectively), 6A (13 and 7%, respectively), 23F (9 and 7%, respectively), and 9V (7 and 7%, respectively) . The theoretical vaccine coverage of the 7-valent conjugate vaccine was 59% for the DCC children and 56% for the NDCC group . Genetic analysis of the pneumococcal isolates revealed 75% clustering among pneumococci isolated from DCC attendees versus 50% among the NDCC children . The average pneumococcal cluster size in the DCC group was 3.8 and 4.6 isolates for two respective sample dates (range, 2 to 13 isolates per cluster), while the average cluster size for the NDCC group was 3.0 (range, 2 to 6 isolates per cluster) . Similar to observations made in other countries, these results indicate a higher risk for horizontal spread of pneumococci in Dutch DCCs than in the general population . This study emphasizes the importance of molecular epidemiological monitoring before, during, and after implementation of pneumococcal conjugate vaccination in national vaccination programs for children. J Clin Microbiol, 2001 Sep, 39(9), 3290 - 5 Genetic and phenotypic features of Streptococcus pyogenes strains isolated in Brazil that harbor new emm sequences; Teixeira LM et al.; In the present study, 37 group A Streptococcus (GAS) strains belonging to 13 new emm sequence types identified among GAS strains randomly isolated in Brazil were characterized by using phenotypic and genotypic methods . The new types were designated st204, st211, st213, st809, st833, st854, st2904, st2911, st2917, st2926, st3757, st3765, and st6735 . All isolates were susceptible to the antimicrobial agents tested, except to tetracycline . They all carried the speB gene, and 94.6% produced detectable SpeB . Most strains belonging to a given emm type had similar or highly related pulsed-field gel electrophoresis profiles that were distinct from profiles of strains of another type . The other characteristics were variable from isolate to isolate, although some associations were consistently found within some emm types . Unlike the other isolates, all type st213 isolates were speA positive and produced SpeA . Strains belonging to st3765 were T6 and opacity factor (OF) negative . Individual isolates within OF-positive emm types were associated with unique sof gene sequence types, while OF-negative isolates were sof negative by PCR . This report provides information on new emm sequence types first detected in GAS isolates from a geographic area not extensively surveyed . Such data can contribute to a better understanding of the local and global dynamics of GAS populations and of the epidemiological aspects of GAS infections occurring in tropical regions. Appl Environ Microbiol, 2001 Sep, 67(9), 3888 - 96 Self-protection against cell wall hydrolysis in Streptococcus milleri NMSCC 061 and analysis of the millericin B operon; Beukes M et al.; Streptococcus milleri NMSCC 061 produces an endopeptidase, millericin B, which hydrolyzes the peptide moiety of susceptible cell wall peptidoglycan . The nucleotide sequence of a 4.9-kb chromosomal region showed three open reading frames (ORFs) and a putative tRNA(Leu) sequence . The three ORFs encode a millericin B preprotein (MilB), a putative immunity protein (MilF), and a putative transporter protein (MilT) . The milB gene encodes a 277-amino-acid preprotein with an 18-amino-acid signal peptide with a consensus IIGG cleavage motif . The predicted protein encoded by milT is homologous to ABC (ATP-binding cassette) transporters of several bacteriocin systems and to proteins implicated in the signal-sequence-independent export of Escherichia coli hemolysin A . These similarities strongly suggest that the milT gene product is involved in the translocation of millericin B . The gene milF encodes a protein of 302 amino acids that shows similarities to the FemA and FemB proteins of Staphylococcus aureus, which are involved in the addition of glycine to a pentapeptide peptidoglycan precursor . Comparisons of the cell wall mucopeptide of S . milleri NMSCC 061(resistant to lysis by millericin B) and S . milleri NMSCC 051(sensitive) showed a single amino acid difference . Serial growth of S . milleri NMSCC 051 in a cell wall minimal medium containing an increased concentration of leucine resulted in the in vivo substitution of leucine for threonine in the mucopeptide of the cell wall . A cell wall variant of S . milleri NMSCC 051 (sensitive) that contained an amino acid substitution (leucine for threonine) within its peptidoglycan cross bridge showed partial susceptibility to millericin B . The putative tRNA(Leu) sequence located upstream of milB may be a cell wall-specific tRNA and could together with the milF protein, play a potential role in the addition of leucine to the pentapeptide peptidoglycan precursor and thereby, contributing to self-protection to millericin B in the producer strain. Scand J Infect Dis, 2001, 33(8), 634 - 7 Culture-negative severe septic shock: indications for streptococcal aetiology based on plasma antibodies and superantigenic activity; Norrby-Teglund A et al.; We present a severe septic shock syndrome patient with negative blood cultures . Acute and convalescent plasma samples from the patient were analysed for anti-streptolysin O titres, superantigen-neutralizing activity and presence of superantigenic activity . The plasma analyses implicated superantigen-producing Streptococcus pyogenes as the causative agent. Structure (Camb), 2001 Apr 4, 9(4), 289 - 97 Crystal structure of Streptococcus mutans pyrophosphatase: a new fold for an old mechanism; Merckel MC et al.; BACKGROUND: Streptococcus mutans pyrophosphatase (Sm-PPase) is a member of a relatively uncommon but widely dispersed sequence family (family II) of inorganic pyrophosphatases . A structure will answer two main questions: is it structurally similar to the family I PPases, and is the mechanism similar? RESULTS: The first family II PPase structure, that of homodimeric Sm-PPase complexed with metal and sulfate ions, has been solved by X-ray crystallography at 2.2 A resolution . The tertiary fold of Sm-PPase consists of a 189 residue alpha/beta N-terminal domain and a 114 residue mixed beta sheet C-terminal domain and bears no resemblance to family I PPase, even though the arrangement of active site ligands and the residues that bind them shows significant similarity . The preference for Mn2+ over Mg2+ in family II PPases is explained by the histidine ligands and bidentate carboxylate coordination . The active site is located at the domain interface . The C-terminal domain is hinged to the N-terminal domain and exists in both closed and open conformations . CONCLUSIONS: The active site similiarities, including a water coordinated to two metal ions, suggest that the family II PPase mechanism is "analogous" (not "homologous") to that of family I PPases . This is a remarkable example of convergent evolution . The large change in C-terminal conformation suggests that domain closure might be the mechanism by which Sm-PPase achieves specificity for pyrophosphate over other polyphosphates. Compend Contin Educ Dent Suppl, 1996, 17(19), S17 - 21 Antibacterial activity of baking soda; Drake D; The antibacterial activity of baking soda (sodium bicarbonate) was assessed using three different experimental approaches . Standard minimum inhibitory concentration analyses revealed substantial inhibitory activity against Streptococcus mutans that was not due to ionic strength or high osmolarity . Short-term exposure assays showed significant killing of bacterial suspensions when baking soda was combined with the detergent sodium dodecylsulfate . Multiple, brief exposures of sucrose-colonized S mutans to baking soda and sodium dodecylsulfate caused statistically significant decreases in numbers of viable cells . Use of oral health care products with high concentrations of baking soda could conceivably result in decreased levels of cariogenic S mutans in saliva and plaque. Arch Intern Med, 2001 Aug 13-27, 161(15), 1866 - 72 Nonsevere community-acquired pneumonia: correlation between cause and severity or comorbidity; Falguera M et al.; BACKGROUND: Community-acquired pneumonia frequently constitutes a nonsevere infection manageable at home . However, for these low-risk episodes, the epidemiological features have not been carefully analyzed . OBJECTIVES: To determine the cause of nonsevere community-acquired pneumonia and to investigate if a correlation exists between cause and severity or comorbidity . METHODS: During a 3-year period, all patients with nonsevere community-acquired pneumonia, according to the Pneumonia Patient Outcome Research Team prognostic classification (patients in groups 1-3), were included in the study . Causes were investigated through the following procedures: cultures of blood, sputum, and pleural fluid; serologic tests; and polymerase chain reaction methods to detect Streptococcus pneumoniae DNA in whole blood or Mycoplasma pneumoniae and Chlamydia pneumoniae DNA in throat swab specimens . RESULTS: Of 317 initially included patients, 247 were eligible for the study . A microbial diagnosis was obtained in 162 patients (66%), and the main pathogens detected were S pneumoniae (69 patients {28%}), M pneumoniae (40 patients {16%}), and C pneumoniae (28 patients {11%}) . For the 58 patients in prognostic group 1, M pneumoniae was the most prevalent cause, and atypical microorganisms constituted 40 (69%) of the isolated agents . In contrast, for patients in prognostic groups 2 and 3, S pneumoniae was the leading agent, and a significant reduction of M pneumoniae cases and a greater presence of other more uncommon pathogens were observed . The existence of comorbid conditions was not a determining factor for particular causes . CONCLUSIONS: Among low-risk patients with community-acquired pneumonia, there was a certain correlation between severity and cause . In contrast, the existence of a comorbidity did not have a predictive causative value. Clin Infect Dis, 2001 Sep 15, 33 Suppl 3, S187 - 92 Antimicrobial use and the emergence of antimicrobial resistance with Streptococcus pneumoniae in the United States; Doern GV; The rapid emergence of resistance to antimicrobial agents by Streptococcus pneumoniae in the United States has been influenced by various factors, including the clonal nature of most resistant strains and the fact that organisms with a multiresistant phenotype have become stably endemic . The ease with which transmission occurs and the fact that humans, especially children, are often colonized asymptomatically in the upper respiratory tract have contributed to the problem . Clearly, the most important factor in the emergence of antimicrobial resistance with S . pneumoniae, however, is the selective pressure of antimicrobial agents . Potency, defined as a product of both antibacterial effect and drug delivery, is a key factor . Generally speaking, the more potent an antimicrobial agent, the less likely it is to select for resistance . This is germane to comparisons of oral agents within specific antimicrobial classes (e.g., beta-lactams, macrolides, and fluoroquinolones) . Within each class, potencies differ . In view of the existence of stably endemic multidrug-resistant S . pneumoniae, given comparable cost, side-effect profile, palatability, convenience of dosing, and accessibility, use of the most potent agent(s) within a particular class is advocated. Clin Microbiol Infect, 2001, 7 Suppl 3, 11 - 7 Structure-activity relationships of ketolides vs . macrolides; Douthwaite S; Since their discovery, the macrolide antimicrobials have proved clinically valuable for the treatment of respiratory tract infections, offering coverage against a broad spectrum of pathogens and excellent tolerability . However, the global increase in macrolide resistance among respiratory pathogens, particularly Streptococcus pneumoniae, threatens their future usefulness . The ketolides, of which telithromycin is the first to reach clinical development, represent a new generation of antimicrobials that have been developed with a view to overcoming the problem of macrolide resistance . Telithromycin is structurally derived from macrolides, and possesses several distinguishing features that are important for its improved microbiological profile . The L-cladinose at position C3 of the miacrolactone ring has been replaced with a keto function . This modification enables telithromycin to bind to its target without tripping the inducible resistance to macrolide-lincosamide-streptograminB (MLS(B)) drugs that many groups of pathogens exhibit . The C6 position has been modified by the addition of a methoxy group . This helps prevent hemiketalization of the C6 position with the 3- and 9-keto groups, thereby conferring excellent acid stability, particularly at gastric pH values . Telithromycin is differentiated from other ketolide compounds by the addition of a large aromatic N-substituted carbamate extension from positions C11/C12 . This carbamate extension improves binding of the drug to its target, the 50S ribosomal subunit, as demonstrated in in vitro experiments . Telithromycin binds to wild-type ribosomes with 10-fold greater affinity than erythromycin A and 6-fold greater affinity than clarithromycin; its affinity for MLS(B)-resistant ribosomes is > 20 times that of both macrolides . The increased ribosomal affinity of telithromycin correlates with its superior potency against Gram-positive cocci both in vitro and in vivo, and is one of the factors determining the drug's activity against MLS(B)-resistant respiratory pathogens. J Biol Chem, 2001 Oct 26, 276(43), 39618 - 28 Epub 2001 Aug 24. Functional analysis of Streptococcus pneumoniae MurM reveals the region responsible for its specificity in the synthesis of branched cell wall peptides; Filipe SR et al.; The recently identified murMN operon of Streptococcus pneumoniae encodes enzymes involved in the synthesis of branched structured muropeptides of the pneumococcal cell wall peptidoglycan . Its inactivation was shown to cause production of a peptidoglycan composed exclusively of linear muropeptides and a virtually complete loss of resistance in penicillin-resistant strains . The studies described in this communication follow up these observations in several directions . The substrate of the MurM-catalyzed reaction (addition of alanine or serine) was identified as the lipid-linked N-acetylglucosamine-muramyl pentapeptide . Different murM alleles from several penicillin-resistant S . pneumoniae strains, each with a characteristic branched peptide pattern, were cloned into pLS578, a pneumococcal plasmid capable of replicating in S . pneumoniae, and transformed into the penicillin-susceptible laboratory strain R36A . All transformants remained penicillin-susceptible; however, their cell wall composition changed in directions corresponding to the muropeptide pattern of the strain from which the murM allele was derived . This suggests that the muropeptide composition of the pneumococcal cell walls is determined by the particular murM allele carried by the cells . A 30-amino acid long sequence within the MurM protein was shown to be the main determinant of the specificity of the reaction (addition of alanine versus serine). J Infect Dis, 2001 Sep 15, 184(6), 794 - 8 Epub 2001 Aug 09. Fluoroquinolone-resistant Streptococcus pneumoniae associated with levofloxacin therapy; Urban C et al.; Fluoroquinolone-resistant cultures of Streptococcus pneumoniae were isolated from 2 patients who were treated for pneumonia with levofloxacin . Nucleotide sequence analysis of bacterial DNA showed that the isolates contained mutations in both parC (DNA topoisomerase IV) and gyrA (DNA gyrase), which were shown previously to confer fluoroquinolone resistance . With the resistant isolates, the MICs for ciprofloxacin, gatifloxacin, grepafloxacin, levofloxacin, and trovafloxacin were above the maximal serum drug concentrations reported for standard dosage regimens . In contrast, the MICs for gemifloxacin and moxifloxacin were below the maximal serum concentrations . Increased effectiveness at blocking the growth of resistant mutants should make gemifloxacin and moxifloxacin less likely to allow the enrichment of mutants within susceptible populations . Additional resistance mutations in the isolates were readily obtained by plating on gemifloxacin- or moxifloxacin-containing agar . Thus, neither compound is expected to halt further accumulation of resistance mutations once mutant enrichment has been initiated by less potent derivatives. J Infect Dis, 2001 Sep 15, 184(6), 789 - 93 Epub 2001 Aug 14. Cross-reactivity of antibodies to type 6B and 6A polysaccharides of Streptococcus pneumoniae, evoked by pneumococcal conjugate vaccines, in infants; Vakevainen M et al.; Streptococcus pneumoniae serotypes 6B and 6A are important causes of infections, yet only 6B is included in current vaccines . It is, therefore, crucial to evaluate whether functional antibodies are produced against both types after vaccination . Concentration and opsonophagocytic activity (OPA) of antibodies to 6B and 6A polysaccharides were determined in serum samples from infants immunized with 3 different pneumococcal conjugate vaccines containing serotype 6B . In all vaccine groups, a significant correlation was found between the anti-6B and -6A antibody concentration and OPA . However, OPA to the vaccine serotype was detectable more commonly than OPA to the cross-reactive type . Furthermore, 5%-15% of the serum samples showed high OPA against the 6B but not the 6A strain . On average, 2-6 times more anti-6B antibodies were needed for 50% opsonophagocytic killing of the type 6A than the type 6B strain . Although pneumococcal type 6B conjugate vaccines elicit antibodies that cross-react with type 6A, not all anti-6B antibodies are functionally cross-reactive. J Infect Dis, 2001 Sep 15, 184(6), 723 - 31 Epub 2001 Aug 10. Streptococcal pyrogenic exotoxin B enhances tissue damage initiated by other Streptococcus pyogenes products; Saouda M et al.; This study compared the pathology and infection pattern of streptococcal pyrogenic exotoxin B-positive (SpeB(+)) and SpeB-negative (SpeB(-)) isogenic variants of an M1 isolate of Streptococcus pyogenes in a mouse skin air sac model . SpeB(+) strains resulted in severe local tissue damage that extended from the epidermis through the subcutaneous layers, whereas isogenic SpeB(-) variants had reduced gross pathology . At the histologic level, differences in necrosis and host responses to each variant were apparent . Injection of purified SpeB alone into a skin air sac failed to induce any significant tissue damage; however, coinjection of the enzyme with either the wild-type or the speB mutant resulted in increased and accelerated tissue necrosis . Surprisingly, coinjection of the enzyme with the spleen-recovered SpeB(-) variant failed to induce a lesion. J Infect Dis, 2001 Sep 15, 184(6), 713 - 22 Epub 2001 Aug 24. Immune-mediated phagocytosis and killing of Streptococcus pneumoniae are associated with direct and bystander macrophage apoptosis; Dockrell DH et al.; Apoptosis of macrophages may be a pathogen-directed mechanism of immune escape or may represent appropriate host response to infection . Human monocyte-derived macrophages (MDMs) from healthy donors (C-MDMs) exhibited low-level constitutive apoptosis, but culture of MDMs with opsonized serotype I Streptococcus pneumoniae (I-MDMs) for 20 h resulted in significantly increased apoptosis . I-MDM apoptosis was associated with phagocytosis of bacteria and intracellular killing that was blocked by the caspase inhibitor z-VAD-fmk but not by Fas-blocking antibody . Paraformaldehyde-fixed I-MDMs induced apoptosis in uninfected syngeneic monocytes at levels greater than those in monocytes incubated alone or incubated with fixed C-MDMs . Apoptosis of syngeneic monocytes was blocked by anti-Fas antibody . The immune response of macrophages to S . pneumoniae includes a novel form of apoptosis that is associated with successful phagocytosis and bacterial killing . This response in vivo may regulate the inflammatory response to infection during a successful host response against S . pneumoniae. Proc Natl Acad Sci U S A, 2001 Aug 28, 98(18), 10416 - 21 Epub 2001 Aug 21. Global differential gene expression in response to growth temperature alteration in group A Streptococcus; Smoot LM et al.; Pathogens are exposed to different temperatures during an infection cycle and must regulate gene expression accordingly . However, the extent to which virulent bacteria alter gene expression in response to temperatures encountered in the host is unknown . Group A Streptococcus (GAS) is a human-specific pathogen that is responsible for illnesses ranging from superficial skin infections and pharyngitis to severe invasive infections such as necrotizing fasciitis and streptococcal toxic shock syndrome . GAS survives and multiplies at different temperatures during human infection . DNA microarray analysis was used to investigate the influence of temperature on global gene expression in a serotype M1 strain grown to exponential phase at 29 degrees C and 37 degrees C . Approximately 9% of genes were differentially expressed by at least 1.5-fold at 29 degrees C relative to 37 degrees C, including genes encoding transporter proteins, proteins involved in iron homeostasis, transcriptional regulators, phage-associated proteins, and proteins with no known homologue . Relatively few known virulence genes were differentially expressed at this threshold . However, transcription of 28 genes encoding proteins with predicted secretion signal sequences was altered, indicating that growth temperature substantially influences the extracellular proteome . TaqMan real-time reverse transcription-PCR assays confirmed the microarray data . We also discovered that transcription of genes encoding hemolysins, and proteins with inferred roles in iron regulation, transport, and homeostasis, was influenced by growth at 40 degrees C . Thus, GAS profoundly alters gene expression in response to temperature . The data delineate the spectrum of temperature-regulated gene expression in an important human pathogen and provide many unforeseen lines of pathogenesis investigation. Infect Dis Obstet Gynecol, 2001, 9(3), 125 - 32 Differences in innate immunologic response to group B streptococcus between colonized and noncolonized women; Smith JM et al.; OBJECTIVE: To evaluate the functional capacity of granulocytes and monocytes from pregnant and nonpregnant women in relation to group B streptococcus (GBS) colonization status . METHODS: Engulfment of fluorescent GBS by peripheral blood phagocytes from GBS-colonized and noncolonized women was measured by flow cytometry . Intracellular superoxiode generated in response to GBS challenge to monocytes and granulocytes enriched from peripheral blood of these women was also measured by flow cytometry, and extracellular superoxide was determined by colorimetric assay . RESULTS: Monocytes and granulocytes from pregnant, GBS-colonized women engulfed significantly greater numbers of GBS than phagocytes from pregnant, noncolonized women . No difference in intracellular superoxide production was detected between any of the groups of women; however, monocytes from pregnant, colonized women released significantly more superoxide into the extracellular milieu than did granulocytes from the same women . No differences in extracellular release of superoxide were observed among noncolonized women whether they were pregnant or not . CONCLUSIONS: Monocytes from pregnant, colonized women engulf more GBS and release more of the superoxide into the extracellular environment, where it is unlikely to be an effective defense mechanism against intracellular bacteria . This suggests that components of the innate immune system that should serve in a protective role may function suboptimally, thereby contributing to the colonization process by GBS. Microsc Res Tech, 2001 Aug 15, 54(4), 254 - 9 Cytochemical study of Streptococcus agalactiae and macrophage interaction; Teixeira CF et al.; Light and electron microscopy were used to analyse the process of interaction of Streptococcus agalactiae (serotypes Ia, III, and V) with resident and activated mouse peritoneal macrophages . Transmission electron microscopy showed that adherence of the S . agalactiae serotype Ia, but not III and V serotypes, to the surface of activated macrophages triggers the respiratory oxidative burst as revealed by the presence of reduced nicotinamide adenine dinucleotide (phosphate) {NAD(P)H}-oxidase in the phagocytic vacuoles . Fusion of macrophage lysosomes with bacteria-containing phagocytic vacuoles was observed in macrophages treated with Lucifer yellow as well as by localization of acid phosphatase for all serotypes . J Bacteriol, 2001 Sep, 183(18), 5436 - 40 Distribution of the SsuDAT1I restriction-modification system among different serotypes of Streptococcus suis; Sekizaki T et al.; The SsuDAT1I restriction-modification (R-M) system, which contains two methyltransferases and two restriction endonucleases with recognition sequence 5'-GATC-3', was first found in a field isolate of Streptococcus suis serotype 2 . Isoschizomers of the R-M system were found in the same locus between purH and purD in a field isolate of serotype 1/2 and the reference strains of serotypes 3, 7, 23, and 26 among 29 strains of different serotypes examined in this study . The R-M gene sequences in serotypes 1/2, 3, 7, and 23 were very similar to those of SsuDAT1I, whereas those in serotype 26 were less similar . These results indicate intraspecies recombination among them and genetic divergence through their evolution. Control Clin Trials, 2001 Aug, 22(4), 438 - 52 Design of a group-randomized Streptococcus pneumoniae vaccine trial; Moulton LH et al.; A group-randomized, double-masked, phase III trial of a Streptococcus pneumoniae conjugate vaccine is being conducted in American Indian populations in the southwestern United States . Approximately 9000 infants will be enrolled in the primary efficacy cohort with vaccine allocation determined by community of residence . The trial is designed to continue until 48 cases of invasive pneumococcal disease due to vaccine serotypes have accumulated . Thirty-eight geographically and socially distinct areas were randomized within blocks formed by population size and geographic location . This design affords the opportunity to capture the effects of herd immunity (indirect effects) by estimating the impact of the vaccine intervention on nonimmunized infants . Group-randomized trials have challenging design and analysis features, many of which are discussed here in the context of the first such trial designed to lead to licensure of a drug or biologic in the United States. Peptides, 2001 Sep, 22(9), 1485 - 9 Adrenomedullin expression in pathogen-challenged oral epithelial cells; Kapas S et al.; Adrenomedullin, a multifunctional peptide, is expressed by many surface epithelial cells and, previously, we have demonstrated that adrenomedullin has antimicrobial activity . The oral cavity contains an epithelium that is permanently colonized by microflora, yet infections in a host are rare . We exposed oral keratinocytes to whole, live cells from four microorganisms commonly isolated from the oral cavity, Porphyromonas gingivalis, Streptococcus mutans, Candida albicans and Eikenella corrodens . There was upregulation of protein and gene expression in these cells in response to bacterial suspensions, but not with the yeast, Candida albicans . We propose there is a potential role for microbial products in enhancing mucosal defense mechanisms and that adrenomedullin participates in the prevention of local infection, thus contributing to host defense mechanisms. Prep Biochem Biotechnol, 2001 Aug, 31(3), 275 - 90 Cleavage and purification of intein fusion proteins using the Streptococcus gordonii spex system; Myscofski DM et al.; A gram-positive bacterial expression vector using Streptococcus gordonii has been developed for expression and secretion, or surface anchoring of heterologous proteins . This system, termed Surface Protein Expression system or SPEX, has been used to express a variety of surface anchored and secreted proteins . In this study, the Mycobacterium xenopi (Mxe) GyrA intein and chitin binding domain from Bacillus circulans chitinase Al were used in conjunction with SPEX to express a fusion protein to facilitate secretion, cleavage, and purification . Streptococcus gordonii was transformed to express a secreted fusion protein consisting of a target protein with a C-terminal intein and chitin-binding domain . Two target proteins, the C-repeat region of the Streptococcus pyogenes M6 protein (M6) and the nuclease A (NucA) enzyme of Staphylococcus aureus, were expressed and tested for intein cleavage . The secreted fusion proteins were purified from culture medium by binding to chitin beads and subjected to reaction conditions to induce intein self-cleavage to release the target protein . The M6 and NucA fusion proteins were shown to bind chitin beads and elute under cleavage reaction conditions . In addition, NucA demonstrated enzyme activity both before and after intein cleavage. Clin Infect Dis, 2001 Sep 15, 33(6), 806 - 14 Epub 2001 Aug 13. Rheumatic fever in the 21st century; Stollerman GH; In the first half of the twentieth century, the group A streptococcus (GAS) was established as the sole etiologic agent of acute rheumatic fever (ARF) . In the century's latter half, the clinical importance of variation in the virulence of strains of GAS has become clearer . Although still obscure, the pathogenesis of ARF requires primary infection of the throat by highly virulent GAS strains . These contain very large hyaluronate capsules and M protein molecules . The latter contain epitopes that are cross-reactive with host tissues and also contain superantigenic toxic moieties . In settings where ARF has become rare, GAS pharyngitis continues to be common, although it is caused by GAS strains of relatively lower virulence . These strains, however, colonize the throat avidly and stubbornly . Molecularly distinct pyoderma strains may cause acute glomerulonephritis, but they are not rheumatogenic, even though they may secondarily colonize and infect the throat . Guidelines for the diagnosis, treatment, and prevention of GAS pharyngitis and ARF are reviewed with particular reference to the prevalence of the latter in the community. Clin Infect Dis, 2001 Sep 15, 33(6), 797 - 805 Epub 2001 Aug 10. Clinical outcomes of bacteremic pneumococcal pneumonia in the era of antibiotic resistance; Moroney JF et al.; Limited data are available about the impact of antimicrobial resistance on clinical outcomes in cases of pneumococcal pneumonia . This was studied in 146 persons hospitalized with invasive pneumonia due to Streptococcus pneumoniae (minimum inhibitory concentration of cefotaxime, > or = .25 microg/mL) who were identified through population-based active surveillance for the period of November 1994 through April 1996 . Compared with matched control subjects who had infection with more-susceptible S . pneumoniae, the proportion of subjects who died or who were admitted to an intensive care unit did not differ significantly . Multivariable analysis showed no significant contribution of antimicrobial resistance to mortality or the requirement for care in an intensive care unit . The ability to detect an effect of antimicrobial resistance on these important outcome measures may have been influenced by aggressive multidrug empirical therapy in this group of hospitalized patients . Factors other than resistance, such as severity of illness at presentation and advance directive status ("do not resuscitate" orders), appear to have a stronger influence on pneumococcal pneumonia outcomes. Eur Respir J, 2001 Jul, 18(1), 196 - 208 Imaging of pneumonia: trends and algorithms; Franquet T; Pneumonia is one of the major infectious diseases responsible for significant morbidity and mortality throughout the world . Imaging plays a crucial role in the detection and management of patients with pneumonia . This review article discusses the different imaging methods used in the diagnosis and management of suspected pulmonary infections . The imaging examination should always begin with conventional radiography . When the results of routine radiography are inconclusive, computed tomography is mandatory . A combination of pattern recognition with knowledge of the clinical setting is the best approach to the pulmonary infectious processes . A specific pattern of involvement can suggest a likely diagnosis in many instances . In acquired immune deficiency syndrome patients, diffuse ground-glass and interstitial infiltrates are most commonly present in Pneumocystis carinii pneumonia whereas in the nonimmunosuppressed patients, a segmental lobar infiltrate is suggestive of a bacterial pneumonia . Round pneumonia is most often encountered in children than adults and is most often caused by Streptococcus pneumoniae . Different combinations of parenchymal and pleural abnormalities may be suggestive for additional diagnoses . When an infectious pulmonary process is suspected, knowledge of the varied radiographic manifestations will narrow the differential diagnosis, helping to direct additional diagnostic measures, and serving as an ideal tool for follow-up examinations. Eur Respir J, 2001 Jul, 18(1), 184 - 95 Pneumococcal vaccination: current and future issues; Ortqvist A; Infection with Streptococcus pneumoniae remains a major global health burden meaning the development of effective vaccines is urgently needed . The current 23-valent polysaccharide vaccine has been shown to prevent pneumococcal pneumonia in immunocompetent young adults, but not in elderly persons . However, in prevention of invasive pneumococcal disease, the vaccine is efficacious in the elderly and may also be effective in some groups of immunocompromised patients . The polysaccharide vaccine is, therefore, recommended in all older (> or = 55-65 yrs of age) adults and in young children (>2 yrs of age) who have a high risk for pneumococcal disease . Revaccination can be safely performed and is recommended 5 yrs after the first dose . In children <2 yrs of age, the new polysaccharide-protein conjugate vaccines, including 7-11 serotypes, seem to be effective in the prevention of invasive disease, severe pneumonia and serotype-specific (and vaccine-related types) otitis media . The low serotype coverage, need for repeated doses, and high price, may decrease the usefulness of the new conjugates . However, the included serotypes correspond to those most often associated with penicillin resistance and vaccination is, therefore, a possible tool in limiting the spread of antibiotic-resistant pneumococci. J Infect Dis, 2001 Oct 1, 184(7), 853 - 60 Epub 2001 Aug 15. Evidence for superantigen involvement in severe group a streptococcal tissue infections; Norrby-Teglund A et al.; Host-pathogen interactions were studied in tissue biopsy samples from patients with severe invasive group A streptococcus (GAS) infections . Skin, subcutaneous tissue, and fascia biopsy samples were divided into clinical grade 1 (no evidence of inflammation {n=7}) or clinical grade 2 (inflamed tissue--erythema and edema including cellulitis, fasciitis, and necrotizing fasciitis {n=24}) . In situ imaging demonstrated significantly higher bacterial load in biopsy samples of higher clinical grade (P<.05), and the bacterial load correlated with the in vivo expression of the superantigen streptococcal pyrogenic exotoxin F (P<.02) . Increased expression of the interleukin-1 cytokines and significantly higher expression of tumor necrosis factor-beta, interferon-gamma, and the homing receptors CC chemokine receptor 5, CD44, and cutaneous lymphocyte-associated antigen (P<.002-.05) were observed in biopsy samples of higher clinical grade . Thus, the cytokine profile at the local site of infection mimics that of a typical superantigen cytokine response . The findings of this study demonstrate a critical role for superantigens and Th1 cytokines in GAS tissue infections. Planta Med, 2001 Aug, 67(6), 561 - 4 Antibacterial activity of Hydrastis canadensis extract and its major isolated alkaloids; Scazzocchio F et al.; The antibacterial activity of extract and isolated major alkaloids (berberine, beta-hydrastine, canadine and canadaline) of Hydrastis canadensis L . (Ranunculaceae) was evaluated against 6 strains of microorganism: Staphylococcus aureus (ATCC 25 993 and ATCC 6538P), Streptococcus sanguis (ATCC 10 556), Escherichia coli (ATCC 25 922), Pseudomonas aeruginosa (ATCC 27 853) . Bactericidal activity was evaluated by contact test by measuring the "killing time" on a low density bacterial inoculum, and bacteriostatic activity in liquid medium by M.I.C . values . The results provide a rational basis for the traditional antibacterial use of Hydrastis canadensis. J Clin Dent, 2002, 13(1), 44 - 9 Bacterial detachment from salivary conditioning films by dentifrice supernates; van der Mei HC et al.; This study compared the detachment by supernates of nine different dentifrices of four oral bacterial strains adhering to a salivary pellicle in a parallel plate flow chamber . Ultra-thin bovine enamel slabs were coated for 1.5 h with human whole saliva . Following buffer rinsing, a bacterial suspension of Streptococcus oralis, Streptococcus sanguis, Streptococcus mutans or Actinomyces naeslundii was perfused through the flow chamber at a shear rate of 30 s-1 for four hours, and the number of adhering bacteria n4h was enumerated by image analysis after buffer rinsing at the same shear rate . Then, a 25 wt%-dentifrice/water supernate was perfused through the flow chamber for four minutes, followed by eight minutes of buffer rinsing and another enumeration of the number of bacteria that had remained adhering nad . Finally, an air-bubble was passed through the flow chamber to mimic the occasionally high detachment forces occurring in the oral cavity, and the adhering bacteria nab were counted again . On average, S . sanguis was the easiest to detach (73% averaged over all dentifrice supernates), while A . naeslundii was the most difficult (22% on average) . The combined detachment of bacteria by dentifrice supernates and air-bubble ranged from a low of 16% to a high of 80% . Dentifrices containing pyrophosphate and polymeric polyphosphate (hexametaphosphate) surface active ingredients appeared to produce the most consistent and strongest desorption effects on plaque bacteria . Factors apparently important to bacterial detachment from pellicle-covered tooth surfaces by dentifrice formulations include the nature of adhesion of bacterial strains and chemical composition of the dentifrice formulations, including pH, surfactant system and the effect of added ingredients (dispersants, metal ions, peroxides, baking soda). Cancer Res, 2001 Aug 15, 61(16), 6151 - 7 Growth-inhibitory effect of a streptococcal antitumor glycoprotein on human epidermoid carcinoma A431 cells: involvement of dephosphorylation of epidermal growth factor receptor; Yoshida J et al.; An antitumor glycoprotein {streptococcal acidic glycoprotein (SAGP)} purified from an extract of Streptococcus pyogenes inhibited the growth of human epidermoid carcinoma A431 cells overexpressing epidermal growth factor receptor (EGFR) in a time- and a concentration-dependent manner . The antiproliferative effect of SAGP was diminished by preincubating the cells with pertussis toxin and by coadministration of sodium orthovanadate, an inhibitor of protein tyrosine phosphatases (PTPases) . Western blot analysis showed that the immunoreactivity of a M(r) 170,000 band of cell lysate to antiphosphotyrosine antibody was reduced by SAGP, and the effect was abolished by sodium orthovanadate . The phosphotyrosine level of the precipitant with anti-EGFR antibody was reduced by SAGP, which was abolished by preincubation with pertussis toxin or by a coadministration with sodium orthovanadate . The PTPase activity transiently increased in the lysate of cells incubated with SAGP and was inhibitable by sodium orthovanadate . Additionally, preincubation of serum-starved A431 cells with SAGP decreased the epidermal growth factor-induced tyrosine phosphorylation of EGFR, and the effect of SAGP was sodium orthovanadate sensitive . These findings indicate that dephosphorylation of the M(r) 170,000 EGFR by activation of PTPase(s) may be responsible in part for the antiproliferative effect of SAGP on A431 cells. FEMS Microbiol Lett, 2001 Aug 7, 202(1), 125 - 7 Age-dependent presence of antibodies in rat dams, capable of conferring protection against group B Streptococcus infection in neonates; Moore JC et al.; A rat model was used to investigate maternal age-dependent resistance on group B Streptococcus (GBS)-induced mortality of the offspring . Offspring from young (first time) or older (repeat litters) dams were challenged with GBS . There was an approximate log difference in the dose of GBS required to cause identical levels of mortality in the two groups . The sera of the dams from both groups were analysed by whole-cell ELISA, and it was demonstrated that sera from the older dams possessed circulating IgG cross-reactive to GBS . Since IgG is transplacentally transferred, we conclude that this is the method of observed protection. APMIS, 2001 Jun, 109(6), 412 - 8 Aerogenous infection of microbiologically defined minipigs with Streptococcus suis serotype 2 . A new model; Madsen LW et al.; Streptococcus suis serotype 2 is the cause of serious infections in animals and humans, but certain aspects of the infection pathogenesis still remain unclear . In this study an experimental model of aerogenous infection and induction of septicemia with S . suis serotype 2 was established in microbiologically defined Gottingen minipigs . Ten animals were exposed to aerosolized S . suis after previous exposure to mild acetic acid in aerosol . Six of the animals were immunosuppressed with prednisolone acetate on different days . All the animals were monitored clinically until euthanasia on days 6 to 13 after exposure . Necropsy was performed and samples were taken for microbiology, histopathology, and immunohistochemistry Three out of four animals immunosuppressed on days 5 to 7 after exposure developed S . suis septicemia, and S . suis could be detected in the tonsil of the soft palate and/or the nasal cavity of all exposed animals . Thus, using the presented model, local as well as systemic infection with S . suis serotype 2 was established in the Gottingen minipig . Since this breed is defined as free of S . suis and a range of other endemic porcine pathogens, the experimental model could prove useful in the study of this infection. Oper Dent, 2001 Jul-Aug, 26(4), 396 - 400 Adherence of plaque components to different restorative materials; Kawai K et al.; This study compared the amount of artificial plaque synthesized in vitro by Streptococcus sobrinus on various dental materials using radioisotopes . In particular, plaque-retaining capacities of new types of ceramics were the focus of this study . Specimens were fabricated from the following materials (one amalgam alloy {Spherical-D}, one casting gold alloy {Casting Gold TYPE I}, one resin composite {Herculite XR} and three ceramics {Vita Celay Blanks, IPS Empress and Dicor MGC}) . The amount of bacteria and glucans adhered on the specimens was measured after incubation for 24 hours at 37 degrees C with radio-labeled cariogenic bacteria and sucrose . This adhesion test was performed using two different surfaces with 600-grit roughness and clinical smoothness . Irrespective of the surface roughness, the least amount of plaque adhered to the ceramics . However, in the case of the resin composite and amalgam, the amount of bacteria and glucan adhesion decreased dramatically by polishing, though there were no statistically different changes in the amount of bacteria and glucans that adhered to the ceramics even after polishing . In general, the amount of adhered bacteria showed almost the same tendency as that of glucans . Although no statistical differences in the amount of bacteria and glucan adhesion were detected among the three ceramics investigated in this study, a lesser amount of bacteria and glucans adhered to them compared to the other materials. J Vet Med Sci, 2001 Jul, 63(7), 723 - 33 Genotypes of pestivirus RNA detected in live virus vaccines for human use; Giangaspero M et al.; Live virus vaccines for human use, 29 monovalent vaccines against measles, mumps, rubella or polio, eight polyvalent vaccines against measles-mumps-rubella and one bacterial polyvalent vaccine against Streptococcus pneumoniae, were tested by reverse transcriptase-nested PCR for the presence of petivirus or pestivirus RNA . Twenty-four samples were selected from European manufacturers, ten were from U.S.A . and four from Japan . Five (13.1%) out of 38 tested samples were positive for pestivirus RNA . Three vaccines (rubella and two measles) were from Europe and two (mumps and rubella) from Japan . The 5'-untranslated genomic region of the contaminant pestivirus RNA were amplified by reverse transcription-PCR and sequenced . Analyses based on primary nucleotide sequence homology and on secondary structures, characteristic to genotypes, revealed that the cDNA sequences belonged to bovine viral diarrhea virus (BVDV) . A cDNA sequence, detected from one measles sample, belonged to BVDV-1b genotype . Pestiviral cDNA detected from the Japanese mumps and rubella vaccine samples, belonged to the BVDV genotypes 1a and 1c, respectively . Analysis on two cDNA sequences detected from measles and rubella vaccine samples from Europe showed their appurtenance to a new genotype, BVDV-1d . These findings indicate that contamination by animal pestivirus may occur in biological products for human use. An Med Interna, 2001 Jun, 18(6), 317 - 8 {Streptococcus suis meningitis}; Geffner Sclarsky DE et al.; Human infection by Streptococcus suis (S . suis) is a zoonosis, with a known occupational risk and clinical presentation mainly as a purulent meningitis with low mortality and frequent hearing loss and ataxia sequela . Less than 150 human cases have been reported since original one thirty years ago . There is a geographical distribution most patients living in northern Europe and south Asia . S . suis disease in human has been reported in two patients in Spain the last years . We present two patients with S . suis meningitis, both were men with occupation related by pork meet, and good outcome . They come at our hospital in a lapse of one month . Both had neurosensorial hearing loss and walking ataxia . One patient had peripheral facial paralysis and diplopia because of paresia of contralateral sixth nerve, with complete resolution at three months . The rare presentation of S . suis meningitis in our country must not forget us to record the working risk at anamnesis. Antimicrob Agents Chemother, 2001 Sep, 45(9), 2436 - 40 Pharmacokinetic and pharmacodynamic parameters for antimicrobial effects of cefotaxime and amoxicillin in an in vitro kinetic model; Gustafsson I et al.; An in vitro kinetic model was used to study the relation between pharmacokinetic and pharmacodynamic (PK-PD) parameters for antimicrobial effect, e.g., the time above MIC (T>MIC), maximum concentration in serum (C(max)), and area under the concentration-time curve (AUC) . Streptococcus pyogenes and Escherichia coli were exposed to cefotaxime, and the activity of amoxicillin against four strains of Streptococcus pneumoniae with different susceptibilities to penicillin was studied . The drug elimination rate varied so that the T>MIC ranged from 20 to 100% during 24 h, while the AUC and/or the initial concentration (C(max)) were kept constant . For S . pyogenes and E . coli, the maximal antimicrobial effect (E(max)) at 24 h occurred when the antimicrobial concentration exceeded the MIC for 50 and 80% of the strains tested, respectively . The penicillin-susceptible pneumococci (MIC, 0.03 mg/liter) and the penicillin-intermediate strain (MIC, 0.25 mg/liter) showed maximal killing by amoxicillin at a T>MIC of 50% . For a strain for which the MIC was 2 mg/liter, C(max) needed to be increased to achieve the E(max) . Under the condition that C(max) was 10 times the MIC, E(max) was obtained at a T>MIC of 60%, indicating that C(max), in addition to T>MIC, may be an important parameter for antimicrobial effect on moderately penicillin-resistant pneumococci . For the strain for which the MIC was 4 mg/liter, the reduction of bacteria varied from -0.4 to -3.6 log(10) CFU/ml at a T>MIC of 100%, despite an initial antimicrobial concentration of 10 times the MIC . Our studies have shown that the in vitro kinetic model is a useful complement to animal models for studying the PK-PD relationship for antimicrobial effect of antibiotics. Antimicrob Agents Chemother, 2001 Sep, 45(9), 2432 - 5 Resistance of Streptococcus pneumoniae to deformylase inhibitors is due to mutations in defB; Margolis P et al.; Resistance to peptide deformylase inhibitors in Escherichia coli or Staphylococcus aureus is due to inactivation of transformylase activity . Knockout experiments in Streptococcus pneumoniae R6x indicate that the transformylase (fmt) and deformylase (defB) genes are essential and that a def paralog (defA) is not . Actinonin-resistant mutants of S . pneumoniae ATCC 49619 harbor mutations in defB but not in fmt . Reintroduction of the mutated defB gene into wild-type S . pneumoniae R6x recreates the resistance phenotype . The altered enzyme displays decreased sensitivity to actinonin. Antimicrob Agents Chemother, 2001 Sep, 45(9), 2427 - 31 In vitro activity of telithromycin against Spanish Streptococcus pneumoniae isolates with characterized macrolide resistance mechanisms; Morosini MI et al.; The susceptibilities to telithromycin of 203 Streptococcus pneumoniae isolates prospectively collected during 1999 and 2000 from 14 different geographical areas in Spain were tested and compared with those to erythromycin A, clindamycin, quinupristin-dalfopristin, penicillin G, cefotaxime, and levofloxacin . Telithromycin was active against 98.9% of isolates (MICs, < or =0.5 microg/ml), with MICs at which 90% of isolates are inhibited being 0.06 microg/ml, irrespective of the resistance genotype . The corresponding values for erythromycin were 61.0% (MICs, < or =0.25 microg/ml) and >64 microg/ml . The erm(B) gene (macrolide-lincosamide-streptogramin B resistance phenotype) was detected in 36.4% (n = 74) of the isolates, which corresponded to 93.6% of erythromycin-intermediate and -resistant isolates, whereas the mef(A) gene (M phenotype {resistance to erythromycin and susceptibility to clindamycin and spiramycin without blunting}) was present in only 2.4% (n = 5) of the isolates . One of the latter isolates also carried erm(B) . Interestingly, in one isolate for which the erythromycin MIC was 2 microg/ml, none of these resistance genes could be detected . Erythromycin MICs for S . pneumoniae erm(B)-positive isolates were higher (range, 0.5 to >64 microg/ml) than those for erm(B)- and mef(A)-negative isolates (range, 0.008 to 2 microg/ml) . The corresponding values for telithromycin were lower for both groups, with ranges of 0.004 to 1 and 0.002 to 0.06 microg/ml, respectively . The erythromycin MIC was high for a large number of erm(B)-positive isolates, but the telithromycin MIC was low for these isolates . These results indicate the potential usefulness of telithromycin for the treatment of infections caused by erythromycin-susceptible and -resistant S . pneumoniae isolates when macrolides are indicated. Diagn Microbiol Infect Dis, 2001 Jul, 40(3), 95 - 102 Infectious etiologies in acute exacerbation of COPD; Lieberman D et al.; Acute exacerbation (AE) is a frequent episode during the prolonged chronic course of chronic obstructive pulmonary disease (COPD), which entails significant morbidity and mortality . The purpose of this study was to determine the frequency distribution of infectious etiologies in these episodes . Two hundred forty hospitalizations for AECOPD were included in a prospective, purely serologically based study . Paired sera were obtained for each of the hospitalizations and were tested using immunofluorescence or EIA methods to identify 13 different pathogens . Only significant changes in antibody titers were considered diagnostic . The mean age ( +/- SD) of the patients was 66.8 +/- 9.0 years and 179 (84%) were males . In 175 (72.9%) hospitalizations at least one infectious etiology was identified . In 117 (48.8%) hospitalizations at least one of 7 viral etiologies was identified . In 72 (30.0%) hospitalizations at least one of the following atypical bacteria was identified: Legionella spp . in 40 (16.7%), Mycoplasma pneumoniae in 34 (14.2%), and Coxiella burnetii in a single hospitalization . In 58 (24.2%) hospitalizations at least one classic bacterial etiology was found: Streptococcus pneumoniae in 48 (20.0%), Hemophilus influenzae in 10 (4.2%) and Moraxella catarrhalis in 9 (3.8%) . More than one etiology was found in 72 (30.0%) hospitalizations . There were no significant differences in the etiologic distribution when the patients were classified by severity of airway obstruction or the clinical type of the exacerbation . We conclude that in most cases of hospitalization due to AECOPD the infectious etiology is viral or atypical bacteria and is classic bacteria in only a minority of cases . More than one etiologic cause can be identified in a third of the cases . The frequency distribution of the etiologies is not associated with the severity of airway obstruction or the clinical type of the exacerbation . The results of our study suggest that atypical bacteria should be covered in antibiotic regimens recommended for AECOPD . This issue should be addressed in future studies. Br J Nutr, 2001 Aug, 86(2), 301 - 6 Pneumococcal pulmonary infection, septicaemia and survival in young zinc-depleted mice; Strand TA et al.; The aim of the present study was to explore whether mice fed a diet low in Zn (2.0 mg Zn/kg diet) for a relatively short period of time were more prone to severe Streptococcus pneumoniae infection than mice fed a normal diet (25 mg elemental Zn/kg) . The Zn-deficient mice were compared with mice in two Zn-adequate control groups; one pair-fed and another with free access to the diet . After 2 weeks feeding, the mice were infected intranasally under anaesthesia with a suspension containing about 10(7) pneumococci . Clinical status was observed every day and blood samples were examined for S . pneumoniae every second day for a week . All infected mice examined carried the infecting strain intranasally . The survival time and time before positive blood culture were significantly shorter in the Zn-depleted group than in the pair-fed Zn-adequate group (hazard ratios 15.6 and 3.2, and respectively) . At the end of the observation period, ten of the twelve mice in the Zn-deficient group were dead while one of twelve and two of twelve were dead in the two Zn-adequate control groups . This study shows that even acutely-induced Zn deficiency dramatically increases the risk of serious pneumococcal infection in mice. Folia Microbiol (Praha), 2001, 46(1), 3 - 10 Kinetic properties of fractions of extracellular NAD+ nucleosidase from Streptococcus pyogenes as an example of host selection by a pathogen: possible role of serum albumin in the organism; Zahradnik FJ; Preparative isoelectric focusing was used to separate free bacterial NAD+ nucleosidase from its complex with a bound host component . Both fractions were characterized by optimum temperature and activation energy of denaturation . The bacterial product is enzymically inactive . The enzymically active structure is formed upon binding to the host component . Only the host organism can provide the suitable, activating structure . The host component in the present system is added to the cultivation medium with a beef heart extract but it can be replaced by serum albumin . The possible role of albumin as a carrier structure for flexible and enzymically inactive peptides is discussed . Different peptides bound to albumin can provide different enzyme activities . The term binary enzyme is coined, referring to a situation where the two enzyme components are coded at genetically distant loci . The pathogen makes use of the carrier structure of albumin type and produces another polypeptide invested with an enzyme activity convenient for the pathogen. Saudi Med J, 2000 May, 21(5), 487 - 9 Delayed presentation of congenital diaphragmatic hernia in association with group B streptococcus infection in a preterm Omani neonate; Manzar S et al.; We present an interesting case of a preterm Omani newborn that had delayed onset of congenital diaphragmatic hernia in association with group B streptococcus infection . The association and the pathogenesis are supported by literature review . The message to follow is that any neonate with prolonged course of streptococcal pneumonia, with unusual course, should be investigated for presence of congenital diaphragmatic hernia. Infect Immun, 2001 Sep, 69(9), 5768 - 76 Fas/Fas ligand system mediates epithelial injury, but not pulmonary host defenses, in response to inhaled bacteria; Matute-Bello G et al.; The Fas/Fas ligand (FasL) system has been implicated in alveolar epithelial cell apoptosis during pulmonary fibrosis and acute respiratory distress syndrome . However, Fas ligation can also lead to cell activation and cytokine production . The goal of this study was to determine the role of the Fas/FasL system in host defenses against Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae . We administered bacteria by aerosolization into the lungs of Fas-deficient (lpr) mice and wild-type (C57BL/6) mice and measured bacterial clearance at 6 and 12 h . One hour prior to euthanasia, the mice received an intraperitoneal injection of human serum albumin (HSA) for alveolar permeability determinations . At all times after bacterial challenges, the lungs of the lpr mice contained similar or lower numbers of bacteria than those of the C57BL/6 mice . Alveolar permeability changes, as determined by bronchoalveolar lavage fluid HSA concentrations, were less severe in the lpr mice 6 h after the challenges . In response to E . coli, the lpr mice had significantly more polymorphonuclear leukocytes (PMN) and macrophage inflammatory protein 2 in the lungs, whereas histopathologic changes were less severe . In contrast, in response to the gram-positive cocci, the lpr animals had similar or lower numbers of PMN . We conclude that the Fas/FasL system contributes to the development of permeability changes and tissue injury during-gram negative bacterial pneumonia . The Fas/FasL system did not have a major role in the clearance of aerosolized bacteria from the lungs at the bacterial doses tested. Infect Immun, 2001 Sep, 69(9), 5456 - 63 Genetic immunization with the region encoding the alpha-helical domain of PspA elicits protective immunity against Streptococcus pneumoniae; Bosarge JR et al.; Pneumococcal surface protein A (PspA) is a pneumococcal virulence factor capable of eliciting protection against pneumococcal infection in mice . Previous studies have demonstrated that the protection is antibody mediated . Here we examined the ability of pspA to elicit a protective immune response following genetic immunization of mice . Mice were immunized by intramuscular injections with a eukaryotic expression vector encoding the alpha-helical domain of PspA/Rx1 . Immunization induced a PspA-specific serum antibody response, and immunized mice survived pneumococcal challenge . Survival and antibody responses occurred in a dose-dependent manner, the highest survival rates being seen with doses of 10 microg or greater . The ability of genetic immunization to elicit cross-protection was demonstrated by the survival of immunized mice challenged with pneumococcal strains differing in capsule and PspA types . Also, immunized mice were protected from intravenous and intratracheal challenges with pneumococci . Similar to the results seen with immunization with PspA, the survival of mice genetically immunized with pspA was antibody mediated . There was no decline in the level of protection 7 months after immunization . These results support the use of genetic immunization to elicit protective immune responses against extracellular pathogens. Infect Immun, 2001 Sep, 69(9), 5430 - 9 Changes in availability of oxygen accentuate differences in capsular polysaccharide expression by phenotypic variants and clinical isolates of Streptococcus pneumoniae; Weiser JN et al.; Most isolates of Streptococcus pneumoniae are mixed populations of transparent (T) and opaque (O) colony phenotypes . Differences in the production of capsular polysaccharide (CPS) between O and T variants were accentuated by changes in the environmental concentration of oxygen . O variants demonstrated a 5.2- to 10.6-fold increase in amounts of CPS under anaerobic compared to atmospheric growth conditions, while CPS production remained low under all conditions for T variants . Increased amounts of CPS in O compared to T pneumococci were associated with increased expression of cps-encoded proteins . The inhibitory effect of oxygen on expression of CPS in O variants correlated with decreased tyrosine phosphorylation of CpsD, a tyrosine kinase and regulator of CPS synthesis . Modulation of CpsD expression and its activity by tyrosine phosphorylation may allow the pneumococcus to adapt to the requirements of both colonization, where decreased CPS allows for adherence, and bacteremia, where increased CPS may be required to escape from opsonic clearance . In patients with invasive infection, paired isolates from the same patient were shown to have predominantly a T colony phenotype without phosphotyrosine on CpsD when cultured from the nasopharynx, and an O phenotype that phosphorylates CpsD in response to oxygen when cultured from the blood . Differences in the availability of oxygen, therefore, may be a key factor in allowing for the selection of distinct phenotypes in these two host environments. Infect Immun, 2001 Sep, 69(9), 5294 - 304 Pulmonary and systemic host response to Streptococcus pneumoniae and Klebsiella pneumoniae bacteremia in normal and immunosuppressed mice; Wang E et al.; Mortality related to bacteremic pneumonia remains high, and the role of sepsis in inflammation, pulmonary injury, and death remains unclear, mostly in leukopenic states . In the present study, the microbiology, histopathology, and host response to Streptococcus pneumoniae and Klebsiella pneumoniae infection were determined in an experimental model of bacteremia in immunocompetent and leukopenic mice . Leukocyte depletion by cyclophosphamide did not impair the early clearance of pneumococci from blood but facilitated growth in lungs . By contrast, klebsiellae rapidly grew in blood of leukopenic mice . These observations suggest that tissue-based phagocytes and circulating leukocytes, respectively, play prominent roles in S . pneumoniae and K . pneumoniae eradication . The kinetics of leukocyte recruitment in lungs during S . pneumoniae bacteremia suggested early strong inflammation in immunocompetent mice that is associated with tumor necrosis factor alpha release and histological disorders, including cell debris and surfactant in alveolar spaces . Leukocyte depletion further stimulated pulmonary capillary leakage both in S . pneumoniae and K . pneumoniae bacteremia, which seemed attributable to bacterial virulence factors . Nitric oxide production did not differ significantly among groups . Leukopenia and low platelet counts characterized the late stage of bacteremia for both strains, but only K . pneumoniae altered renal function . Understanding the pathogenesis of bacteremia will help establish beneficial therapies for both sepsis and pneumonia. Microb Pathog, 2001 Sep, 31(3), 109 - 14 Complementation of a speA negative Streptococcus pyogenes with speA: effects on virulence and production of streptococcal pyrogenic exotoxin A; Unnikrishnan M et al.; We have shown previously that an isogenic SPEA-negative Streptococcus pyogenes strain did not attenuate virulence in a murine model of necrotizing fasciitis . The aim of this study was to confirm that streptococcal pyrogenic exotoxin A (SPEA) is not crucial for streptococcal invasiveness in murine invasive infection . The SPEA-negative S . pyogenes (H326) was complemented with speA extra-chromosomally to create strain H361 which produced 2.2-fold more SPEA compared with the parental speA(+)wild-type (H305) . The growth phase-regulated expression of SPEA in vitro was unaffected in this strain . Complementation with speA resulted in reduced virulence and bacterial counts in invasive murine infection . SPEA production was quantitated from muscle tissue of infected mice . However, H361 did not produce more SPEA than H305 in vivo . We conclude that SPEA does not play a key role in invasive murine streptococcal infection . Drug Resist Updat, 1999 Dec, 2(6), 347 - 350 10th Conference on DNA Topoisomerases in therapy; Giaccone G et al.; The 10th Conference on DNA Topoisomerases in Therapy 6-8 October 1999 in Amsterdam, The Netherlands) covered basic research on DNA topoisomerases and aspects of topoisomerase-directed therapy . The understanding of basic aspects of enzyme functions and structures was discussed throughout the meeting, as this knowledge is fundamental to further developments of new and more effective therapeutic approaches . Several new crystal structures were presented, and implications for function and interaction with DNA and drugs were discussed . Knock-out mice for various topoisomerase genes have been produced and genes have been shown to differ in importance for development and survival . The interaction of topoisomerases with other proteins involved in DNA metabolism, chromosome stability and physiology were discussed . The main focus for cancer therapy was on camptothecins (CPT) and related compounds stabilizing covalent DNA-intermediates of topoisomerase I . Reports on recent clinical trials of first-generation, water-soluble CPT-analogs (topotecan and irinotecan) confirmed earlier findings of activity in several solid tumors and hematological malignancies . Improvements in efficacy and toxicity profiles are being sought in orally absorbable compounds and other drug formulations (e.g . in liposomes) . Several new CPT-analogs at preclinical stages of development might also provide a greater stability of the lactone ring, higher DNA-binding affinity, and improved water solubility . New drugs have also been developed from a number of new non-CPT compounds, which inhibit the activity of DNA-topoisomerases but do not stabilize the DNA-linked form of the enzymes . Another focus of the meeting was on therapeutic targeting of microbial DNA topoisomerases . The antibiotic potential of the quinolones has been extended to gram-positive pathogens, particularly Streptococcus pneumoniae . The cloning and biochemical characterization of the DNA-topoisomerases of eukaryotic parasites such as Plasmodium falciparum or Candida albicans have been completed and the search for specific inhibitors targeting these enzymes is under way . Am J Vet Res, 2001 Aug, 62(8), 1262 - 5 Characteristics and management practices associated with milk production in dairy herds in Ohio enrolled in official Dairy Herd Improvement Association programs; Khaitsa ML et al.; OBJECTIVE: To determine herd characteristics and management practices associated with milk production in dairy herds enrolled in official Dairy Herd Improvement Association (DHIA) programs in Ohio . SAMPLE POPULATION: 186 dairy farms in Ohio . PROCEDURE: All herds in official DHIA programs in 9 counties were invited to participate . Information regarding herd characteristics and management practices was obtained, using a standardized questionnaire . Bulk-tank milk samples were obtained for bacteriologic culture . Official DHIA test-day records were obtained, and associations were identified, using multivariable ANOVA procedures . RESULTS: Of 479 eligible producers, 186 (39%) participated, and consecutive bulk-tank milk samples were available for culture from 172 (36%) . Streptococcus agalactiae and Mycoplasma spp were not recovered from bulk-tank milk samples, but Staphylococcus aureus was recovered from 64 (37%) herds . Mean (+/- SD) number of lactating cows in participating herds was 97+/-66, with 123 (66%) herds milking < 100 cows . The RHA was significantly associated with number of cows in milk, estimated percentage of herd detected in estrus, reported annual percentage of heifer calves born alive that died before 8 weeks old, percentage days in milk, use of bovine somatotropin during the preceding 2 years, and sex of the person completing the questionnaire . CONCLUSIONS AND CLINICAL RELEVANCE: In this study, the strongest indicator of milk production was number of cows in milk . However, merely adding cows to a herd should not be considered to guarantee increased milk production, because other management traits could be confounded with increased number of cows in a herd. Intensive Care Med, 2001 Jun, 27(6), 965 - 9 Elevation of troponin I in sepsis and septic shock; Ammann P et al.; OBJECTIVE: To detect myocardial damage in severe systemic inflammation by cTnI measurements in patients without acute coronary syndromes . DESIGN: Prospective case control study . SETTING: Tertiary referral center . PARTICIPANTS: Twenty patients with sepsis, septic shock, and systemic inflammatory response syndrome (SIRS) were examined and compared to controls without coronary artery disease or myocarditis . MEASUREMENTS AND RESULTS: cTnI levels were assessed in patients with SIRS, sepsis, and septic shock . Eight patients (two female/six male) suffered from septic shock, nine (three female/six male) from sepsis without shock, and three (three male) from SIRS . Seventeen patients (85%) showed elevated cTnI (median 0.57 microg/l; 0.17-15.4), whereas no patient in the control group showed elevated cTnI (P < 0.0001) . Six patients (30%),--three with septic shock and three with sepsis--died during hospitalization, five of them with elevated cTnI . Four out of five autopsies showed normal coronary arteries . Coronary angiography, autopsy, and stress echocardiography ruled out significant coronary artery disease in ten cTnI-positive patients (59%) . In 41 % of cTnI-positive patients, Streptococcus pneumoniae could be cultured, whereas no cTnI-negative or control patient showed signs of infection due to S . pneumoniae . CONCLUSION: Cardiac troponin I was elevated in 85% of patients with sepsis, septic shock or SIRS in our study . A high percentage showed infection caused by S . pneumoniae . In what way microorganisms cause cTnI elevations is not yet understood. Kansenshogaku Zasshi, 2001 Jun, 75(6), 464 - 8 {Clinical and bacteriological features of 12 cases of liver abscess caused by Streptococcus milleri group}; Kanamori S et al.; We described the clinical and bacteriological features of 12 cases of liver abscess caused by Streptococcus milleri group (SMG) during a 6-year period from 1993 to 1998 . The gender was 11 males and 1 female with their ages ranging from 39 to 76 years old (mean: 53.4) . The common symptoms were fever (100%), abdominal pain (67%), and appetite loss (58%) . Nine cases had underlying diseases such as carcinomas and diabetes mellitus . Predominant causes of the liver abscess were cryptogenic (42%) and biliary tract disease (33%) . Three patients died of an exacerbation of the carcinoma . Eight cases (67%) was single infection of SMG and no mixed infection with anaerobes . No strains isolated in this series showed resistance against penicillin G and ampicillin . SMG was highly isolated from the blood culture in eight of the 11 cases (73%) . Liver abscess should be taken into consideration as one of the causes of SMG septicemia. J Infect Dis, 2001 Sep 1, 184(5), 577 - 81 Epub 2001 Jul 30. Do antibodies to pneumococcal surface adhesin a prevent pneumococcal involvement in acute otitis media? Rapola S, Kilpi T, Lahdenkari M, Takala AK, Makela PH, Kayhty H. Antibodies to the pneumococcal (Pnc) surface protein PsaA are immunogenic and protective in experimental animal models, but their role in protection from Pnc disease in humans is not known . In the present study, the ability of antibodies to PsaA to prevent the progression of Pnc carriage to Pnc acute otitis media (Pnc AOM) was evaluated . Antibodies to PsaA were measured in acute-phase serum samples of children with AOM and with Streptococcus pneumoniae cultured from the nasopharynx . The risk of Pnc AOM was evaluated by a logistic regression model with anti-PsaA concentration as the predictive variable . Higher concentrations of antibodies to PsaA were associated with lower risk of the Pnc nasopharyngeal carriage progression to Pnc AOM . This was true in children 9-24 months old (odds ratio {OR}, 0.49; 95% confidence interval {CI}, 0.31-0.78) but not in children <9 months old (OR, 0.81; 95% CI, 0.48-1.35). J Infect Dis, 2001 Sep 1, 184(5), 569 - 76 Epub 2001 Aug 02. Natural development of antibodies to pneumococcal capsular polysaccharides depends on the serotype: association with pneumococcal carriage and acute otitis media in young children; Soininen A et al.; To study the natural development of antibodies to pneumococcal capsular polysaccharides of types 1, 6B, 11A, 14, 19F, and 23F and its association with pneumococcal carriage and acute otitis media (AOM), 329 children were followed-up prospectively during their first 2 years of life . Nasopharyngeal carriage was determined by cultures of nasopharyngeal swab samples, and etiology of AOM was determined by cultures of middle ear fluid . Antibodies were measured in serum samples collected at 6, 12, 18, and 24 months by EIA . Antibodies increased modestly but significantly with age . Contact with serotypes 11A and 14 was associated with increased antibody concentration as early as age 6 months . Children with contact with serotypes 6B, 19F, and 23F had antibody levels similar to those in children without contact . Antibodies increased modestly, even in children without known contact with Streptococcus pneumoniae and in children with contact with heterologous serotypes . Antibody concentrations were equal after carriage or AOM. Braz J Infect Dis, 2001 Apr, 5(2), 50 - 2 Epub 2001 Aug 03. In vitro activity of fluoroquinolones (gatifloxacin, levofloxacin and trovafloxacin) and seven other antibiotics against Streptococcus pneumoniae; Nicodemo AC et al.; In recent years, the level of resistance of S . pneumoniae to beta-lactam and/or macrolides has increased around the world including some countries in South America . Because of this resistance, it is necessary to test the therapeutic alternatives for treating this pathogen, including the newer quinolones . This study was carried out in order to compare the in vitro activity of fluoroquinolones gatifloxacin, levofloxacin and trovafloxacin, to penicillin G, amoxicillin, amoxicillin-clavulanate, cufuroxime sodium, ceftriaxone, azithromycin and clarithromycin, against 300 strains of S . pneumoniae . Of the 300 samples tested, 18.6% were not susceptible to penicillin (56 strains) and 7% (21 strains) were resistant to the second generation cephalosporin . Among the macrolides, resistance ranged from 6.7% for clarithromycin to 29.6% for azithromycin . Susceptibility to the newer quinolones was 100% including the 56 strains not susceptible to penicillin . Among the 10 antibiotics evaluated, the fluoroquinolones gatifloxacin, levofloxacin, and trovafloxacin displayed high levels of in vitro activity against S . pneumoniae. J Intern Med, 2001 Aug, 250(2), 174 - 9 A case of toxic shock-like syndrome presenting with serious hypoproteinaemia because of a protein-losing gastroenteropathy; Tasaki K et al.; A 37-year-old man was admitted to our hospital because of toxic shock-like syndrome (TSLS) induced by Streptococcus pyogenes . After the pathogenic bacteria had been eradicated, serious diarrhoea appeared and a protein-losing gastroenteropathy developed . An immunohistochemical study of the biopsy specimens of both small and large intestines revealed the infiltration of T-lymphocytes, predominantly CD8+ cells, into the lamina propria of affected mucosa, villus atrophy and crypt hyperplasia . Considering these histological findings, some immunological mechanism which lead the activation of cytotoxic T-lymphocytes may play an important role in the pathogenesis of this rare intestinal manifestation of TSLS. Contemp Top Lab Anim Sci, 2000 Jul, 39(4), 43 - 6 Endocarditis and meningitis caused by Streptococcus suis after cardiac surgery in a sheep; Wilson RP et al.; A female Dorset-cross sheep developed labored respirations and was anorexic, weak, ataxic and febrile (42.0 degrees C) 15 days after implantation of a cardiac device . Clinical pathologic evaluation identified a mild leukocytosis (11,800 cells/microl), neutrophilia (8,969 cells/microl), and hypokalemia (2.9 mmol/L) . Despite intensive therapy, the animal was euthanized 7 days later . Necropsy of the sheep revealed meningitis and vegetative endocarditis associated with the cardiac device . Blood cultures collected antemortem and tissue cultures collected at necropsy yielded Streptococcus suis . Histopathology confirmed the diagnosis of bacterial valvular endocarditis and meningoencephalitis . A variety of environmental and host factors are proposed as contributing to the infection, and the zoonotic potential of S . suis is discussed. Clin Infect Dis, 2001 Sep 1, 33(5), 740 - 4 Epub 2001 Aug 06. Necrotizing fasciitis due to Streptococcus pneumoniae after intramuscular injection of nonsteroidal anti-inflammatory drugs: report of 2 cases and review; Frick S et al.; Two cases of pneumococcal necrotizing fasciitis (NF) occurred after intramuscular injections of nonsteroidal anti-inflammatory drugs; another 5 cases reported in the literature fulfilled the criteria for NF involving Streptococcus pneumoniae . Conditions associated with alterations of immune function could be identified in 6 of the 7 cases; 2 patients died despite surgical and antimicrobial treatment. Clin Infect Dis, 2001 Sep 1, 33(5), 662 - 75 Epub 2001 Jul 30. Rethinking recommendations for use of pneumococcal vaccines in adults; Whitney CG et al.; Streptococcus pneumoniae remains a major cause of disease worldwide; the emergence of antibiotic-resistant strains emphasizes the importance of disease prevention by use of vaccines . Recent studies have provided information that is useful for the evaluation of current vaccine recommendations . Recommendations target most people who are at high risk for invasive pneumococcal disease . However, higher risk has also been identified for African Americans and smokers, but these groups are not specifically targeted by current recommendations . The vaccine is effective against invasive disease in immunocompetent people, although studies in immunocompromised subjects have found few subgroups in which the vaccine appears to be effective . Questions with regard to optimal timing and indications for revaccination remain a challenge, because the duration of protection and effectiveness of revaccination remain unknown . New pneumococcal vaccines appear promising but will need to be tested against the performance of the polysaccharide vaccine . Improving delivery of the currently available pneumococcal polysaccharide vaccine to adults who will benefit should be a high priority. Arch Intern Med, 2001 Jul 23, 161(14), 1742 - 8 Pneumococcal peritonitis in adult patients: report of 64 cases with special reference to emergence of antibiotic resistance; Capdevila O et al.; BACKGROUND: Few data are available regarding pneumococcal peritonitis . We studied the clinical characteristics of intra-abdominal infections caused by Streptococcus pneumoniae and its prognosis in relation to antibiotic resistance . METHODS: We reviewed all cases of culture-proved pneumococcal peritonitis . Patients with liver cirrhosis and primary pneumococcal peritonitis were compared with patients with Escherichia coli peritonitis . RESULTS: Between January 1, 1979, and December 31, 1998, we identified 45 cases of primary pneumococcal peritonitis in patients with cirrhosis and 19 cases of secondary (or tertiary) pneumococcal peritonitis . Patients with cirrhosis and primary pneumococcal peritonitis vs those with primary E coli peritonitis had more frequent community-acquired infection, 73% vs 47%; pneumonia, 36% vs 2%; and bacteremia, 76% vs 33%; and higher attributable mortality (early mortality), 27% vs 9% (P<.05 for all) . Secondary (or tertiary) pneumococcal peritonitis was associated with upper or lower gastrointestinal tract diseases; in most cases, the infection appeared after surgery . A hematogenous spread of S pneumoniae from a respiratory tract infection might be the most important origin of peritonitis; also, S pneumoniae might directly reach the gastrointestinal tract favored by endoscopic procedures or hypochlorhydria . There was an increased prevalence of penicillin and cephalosporin resistance up to 30.7% and 17.0%, respectively, although it was not associated with increased mortality rates . CONCLUSIONS: Primary pneumococcal peritonitis in patients with cirrhosis more often spread hematogenously from the respiratory tract and was associated with early mortality . In secondary (and tertiary) pneumococcal peritonitis, a transient gastrointestinal tract colonization and inoculation during surgery might be the most important mechanisms . Current levels of resistance were not associated with increased mortality rates. Southeast Asian J Trop Med Public Health, 2001 Mar, 32(1), 126 - 31 Invasive pneumococcal infection in children; Yu CH et al.; Streptococcus pneumoniae infections can involve multiple organs and cause high mortality and morbidity . In this retrospective study, we reviewed patients with invasive pneumococcal infection in the pediatric department of a teaching hospital in Taipei . From 1984 to 1998, 81 children with invasive pneumococcal infection were hospitalized . Twenty-eight patients had meningitis, 27 had pneumonia with pleural effusion, 60 had sepsis, and 4 had arthritis . Thirty-eight patients had more than one site of infection . Most of our patients (81.7%) were below 5 years of age . Pneumococcal infections were more common from October to March . Eight patients had a history of trauma that correlated with the site of infection . Thirteen patients (16.0%) expired and 20 (24.7%) had severe sequelae . Multi-regression analysis found that meningitis and complications were independent variables that affected the outcome . The percentage of penicillin-resistant strains increased beginning in 1990 and accounted for about four-fifths of the infections in the final 2 years of the study . Since invasive pneumococcal infections in children may have a poor prognosis and penicillin-resistant strains have become increasingly common, early and adequate antibiotic therapy should be given as soon as possible. Acta Neuropathol (Berl), 2001 May, 101(5), 499 - 508 A mouse model of Streptococcus pneumoniae meningitis mimicking several features of human disease; Gerber J et al.; The course of bacterial titers, meningeal inflammation, behavioral abnormalities, and neuronal damage was studied in a mouse model of Streptococcus pneumoniae meningitis . At 24 h after injection of 10(4) colony-forming units (CFU) S . pneumoniae into the right forebrain, infected mice became severely lethargic . Bacterial titers in cerebrospinal fluid and cerebellum rose to 10(9) CFU/ml, with strong granulocyte invasion into the meninges and neuronal necroses in the neocortex, striatum and hippocampal formation . Meningeal inflammation and neuronal damage in intercellular cell adhesion molecule-1- and macrophage colony-stimulating factor-deficient mice was similar to that in wild-type littermates . Untreated, the infection was fatal . Wild-type mice treated earlier than 24 h after infection with ceftriaxone (2 mg every 12 h for 3 days) survived without apparent behavioral abnormalities . Delay of treatment beyond 30 h led to the death of more than 50% of the infected mice . This mouse model is suitable for therapeutic studies and for the investigation of inflammation in knockout mice . The neuronal damage resembles morphological abnormalities observed in humans. Acta Clin Belg, 2000 Nov-Dec, 55(6), 312 - 22 In vitro study on the antimicrobial activity of various antibiotics against clinical isolates of Streptococcus pneumoniae from Belgium collected during winter 1998-1999; Vanhoof R et al.; A total of 205 isolates of Streptococcus pneumoniae obtained from 10 different centres were included in this study . The susceptibilities to penicillin, ampicillin, amoxicillin, amoxicillin/clavulanic acid, cefaclor, cefuroxime, cefotaxime, imipenem, ciprofloxacin, gemifloxacin, grepafloxacin, levofloxacin, trovafloxacin, erythromycin, clarithromycin, miocamycin, clindamycin and tetracycline were determined by a microdilution technique following NCCLS recommendations . Decreased susceptibility to penicillin was 16.1% {6.8% intermediate (0.12-1 microgram/mL) and 9.3% high-level (> or = 2 micrograms/mL)}, cefotaxime insusceptibility (> or = 1 microgram/mL) 12.7%, ciprofloxacine insusceptibility (> or = 2 micrograms/mL) 15.6% with 1.5% of high level resistance (> or = 4 micrograms/mL), erythromycin insusceptibility (> or = 0.5 microgram/mL) 36.1% and tetracycline insusceptibility (> or = 4 micrograms/mL) 22.9% . Decreased susceptibility to cefotaxime was found in 78.8% of the penicillin-insusceptible isolates . No decreased susceptibility was found for gemifloxacin (> or = 0.5 microgram/mL) and trovafloxacin (> or = 1 microgram/mL) . Compared to the 1996-1997 surveillance, penicillin, cefotaxime and erythromycin insusceptibility rose by 3.8%, 5.2% and 5.0% respectively, while tetracycline insusceptibility decreased with 8.2% . MICs of all beta-lactams rose with those of penicillin for penicillin-insusceptible isolates . Amoxicillin +/- clavulanate, cefotaxime and imipenem were generally 1, 1 and 5 doubling dilutions respectively more potent than penicillin on these isolates . Penicillin, ampicillin and cefuroxime were equally active while cefaclor was generally 5 dilutions less potent . Most penicillin-insusceptible isolates remained fully susceptible to amoxicillin +/- clavulanate and imipenem . The penicillin-insusceptible isolates were 36.4%, 27.3% and 3.0% co-insusceptible to erythromycin, erythromycin plus tetracycline and tetracycline respectively . A subpopulation of 52 isolates obtained from children aged < or = 3 years was also studied . Compared to the other isolates we found a statistically significant increase in insusceptibility for penicillin, cefaclor, cefuroxime, erythromycin, clarithromycin and tetracycline while a significant decrease was found for ciprofloxacin. J Biomed Mater Res, 2001 Nov, 57(2), 248 - 57 Poly(L-lactide)acid/alginate composite membranes for guided tissue regeneration; Milella E et al.; The barrier membranes for guided tissue regeneration (GTR) to treat bone defects have to satisfy the criteria of biocompatibility, cell-occlusiveness, space-making, tissue integration and clinical manageability . In this study a system constituted of a poly(L-lactide) acid (PLLA) asymmetric membrane combined with an alginate film was prepared . The PLLA membrane functions to both support the alginate film and separate the soft tissue; the alginate film is intended to act as potential vehicle for the growth factors to promote osteogenesis . The structural, morphological, and mechanical properties of the bilamellar membrane and its stability in culture medium were evaluated . Moreover, the feasibility of using the alginate membranes as controlled-release delivery vehicles of TGF-beta was monitored . Finally, the bacterial adhesion and permeability of Streptococcus mutans, selected for the high adhesive affinity, were monitored . The results showed that the surfaces of the alginate side, to be used in contact with the bone defect, were rougher than PLLA ones . When in contact with complete culture medium, the PLLA-alginate membrane retained its mechanical and structural properties for more than 100 days . Then, the degradation processes occurred but the membrane continued to be stable and manageable for 6 months . Growth factors such as TGF-beta can be incorporated into alginate membranes functioning as drug delivery vehicle, and retain the biological activity when tested in an in vitro model system . The obtained membrane acted as a barrier to the passage of S . mutans bacteria and showed to promote a lower bacterial adhesion with respect to commercial GTR membranes . Pediatrics . 2001 Aug;108(2):E23. Serotype prevalence of occult pneumococcal bacteremia; Alpern ER et al.; OBJECTIVE: The licensure and use of a pneumococcal conjugate vaccine that is immunogenic in children who are younger than 2 years may affect the epidemiology of occult bacteremia . This study was conducted to determine the serotype prevalence of Streptococcus pneumoniae isolates from children with occult bacteremia and to document the proportion that would be covered by the recently licensed heptavalent pneumococcal conjugate vaccine . METHODS: A cohort of 5901 children who were 2 to 24 months of age and had a temperature of >/=39.0 degrees C evaluated with a blood culture at an urban tertiary care children's hospital emergency department was studied to determine the prevalence of S pneumoniae serotypes . Patients were excluded if their immune system was suppressed, they had a diagnosis of a focal infection, they were evaluated by lumbar puncture, they were admitted to the hospital, or they died during initial evaluation . Blood cultures were inoculated into pediatric blood culture bottles and processed using an automated carbon dioxide monitoring system . All pneumococcal isolates were serotyped on the basis of capsular swelling with type-specific antisera (Quellung reaction) . RESULTS: The study population consisted of 5901 patients . The overall rate of occult bacteremia was 1.9% (95% confidence interval {CI}: 1.5-2.3) . S pneumoniae accounted for 92 of 111 isolates (82.9%; 95% CI: 74.6-89.4) in children with occult bacteremia . Eight pneumococcal serotypes were represented: 6A (2%), 9V (6%), 19F (6%), 18C (8%), 4 (9%), 6B (13%), 23F (15%), and 14 (42%) . Serotypes 14, 6B, and 23F accounted for 69.3% (95% CI: 58.6-78.7) of typed isolates . In the cohort, 97.7% (95% CI: 92-99.7) of isolated serotypes are represented in the newly licensed heptavalent pneumococcal conjugate vaccine . The single isolated serotype that would not have been covered by the currently licensed heptavalent pneumococcal conjugate vaccine was 6A . CONCLUSIONS: S pneumoniae accounts for the vast majority of bacterial pathogens in children with occult bacteremia . As indicated by the results of this study, the heptavalent pneumococcal conjugate vaccine may prevent the majority of occult pneumococcal bacteremia episodes . The 2 cases of bacteremia with a serotype that would not have been included in the vaccine both were due to serotype 6A . It has been noted that there is potential nonvaccine serotype and subgroup cross-protection (6A from 6B) afforded to children who are immunized with the heptavalent vaccine . The high potential efficacy of the heptavalent pneumococcal conjugate vaccine for strains that cause occult bacteremia in our population may have a profound effect on the treatment of children with fever without a source . There has been an alarming and rapid emergence of antibiotic-resistant pneumococcal strains . Less pressure to use broad-spectrum antibiotics, which in turn causes further antibiotic resistance, should result . Laboratory testing and hospitalization also should be reduced . The prevalence rates determined by this study may be used as baseline data for comparison of serotype rates of occult pneumococcal bacteremia after widespread use of the heptavalent vaccine. Farmaco, 2001 May-Jul, 56(5-7), 417 - 20 Isolation of praeruptorins A and B from Peucedanum praeruptorum Dunn . and their general pharmacological evaluation in comparison with extracts of the drug; Lu M et al.; The root of Peucedanum praeruptorum Dunn . was extracted with solvents at different polarity obtaining three chemical fractions: aqueous (H2O), n-butanol (BuOH) and ethyl acetate (AcOEt) . From AcOEt praeruptorins A and B were isolated by column chromatography on silica gel, using toluene/ethyl acetate as eluent, and identified by 1H and 13C NMR analysis . The extracts and the praeruptorins were tested for gross behavioural effects and acute toxicity in mice; the cytotoxicity on Artemia salina Leach and the antimicrobial activity were also evaluated . None of the tested substances evoked behavioural effects or acute toxicity after oral administration in mice; delayed mortality was observed with AcOEt and praeruptorin A only after intraperitoneal administration of high doses (1 g/kg) . In Artemia salina test AcOEt, and praeruptorins A and B had LC50 values of 40.2, 121.2 and 34.5 microg/ml, respectively . AcOEt and praeruptorin A showed antimicrobial activity on Streptococcus agalactiae; their MIC values were 250 and 100 microg/ml, respectively. J Antimicrob Chemother, 2001 Aug, 48(2), 279 - 81 In vitro activity of cefditoren against clinical isolates of penicillin-susceptible and penicillin-intermediate strains of Streptococcus pneumoniae isolated in Germany, 1992-1998; Reinert RR et al.; This study investigates the susceptibility to cefditoren of penicillin-susceptible strains of invasive Streptococcus pneumoniae (n = 312) and of penicillin-intermediate strains of S . pneumoniae (n = 30) isolated mainly from patients with respiratory tract infections . The MIC(90)s of penicillin-susceptible and -intermediate isolates were as follows: cefditoren, < or =0.06 and 1 mg/L; penicillin G, < or =0.06 and 0.5 mg/L . Cefditoren showed the highest activity against the penicillin-intermediate strains investigated compared with the other beta-lactam antibiotics and is therefore considered to be a promising agent for the treatment of infections caused by pneumococci with reduced penicillin susceptibility. J Antimicrob Chemother, 2001 Aug, 48(2), 195 - 201 The inhibition and selectivity of bacterial topoisomerases by BMS-284756 and its analogues; Lawrence LE et al.; Analogues of BMS-284756, a novel des-F(6)-quinolone, were synthesized and evaluated in order to determine the effects of modification of substituents on in vitro target inhibition . BMS-340281 (stereoisomer of BMS-284756), BMS-340280 (C-6 fluorinated analogue of BMS-284756), BMS-340278 (C-8-H derivative), BMS-433366 (C-8 methoxy analogue) and fluoroquinolone comparators were evaluated for antibacterial activity . The MICs of BMS-284756 were generally found to be within two-fold of the MICs of BMS-284756 analogues against a panel of Gram-positive and -negative organisms . BMS-284756 had MICs of 0.03-0.125 mg/L against Streptococcus pneumoniae strains with GyrA and ParC mutations, and was the most active quinolone . BMS-284756 and its analogues had similar activity compared with ciprofloxacin and moxifloxacin against topoisomerase IV decatenation, but were three times more active than levofloxacin . The IC(50) of BMS-284756 for human topoisomerase II (hTopo II) was 3000 times higher than its IC(50) for DNA gyrase, and no whole-cell cytotoxicity was noted . Two analogues, BMS-340280 and BMS-340278, demonstrated moderate inhibition against hTopo II and cytotoxicity in the cellular assay . BMS-284756 demonstrated greater Gram-positive antibacterial activity and similar inhibition of targets compared with other fluoroquinolones, and more favourable selectivity compared with the other BMS-284756 analogues. Acta Orthop Scand, 2001 Jun, 72(3), 237 - 40 Effective bactericidal activity of tobramycin and vancomycin eluted from acrylic bone cement; Gonzalez Della Valle A et al.; We studied the bioactivity of vancomycin and tobramycin eluted from methylmethacrylate bone cement . Aliquots of the drainage were obtained at 1, 6, 12 and 24 hours following total hip prosthetic implantation with vancomycin-tobramycin-loaded cement in 3 patients . The samples were analyzed with fluorescence polarization immunoassay and bioassay, using group B streptococcus for vancomycin and Escherichia coli for tobramycin . These bacteria were selected due to the effectiveness of vancomycin and poor effectiveness of tobramycin against group B streptococcus and conversely with E . coli . The immunodetection of vancomycin averaged 14 (1 hour), 9 (6 hours), 10 (12 hours) and 11 microg/mL (24 hours) . The bioassay averaged 47, 36, 79 and 41 microg/mL (p = 0.03) . The immunodetection of tobramycin averaged 43, 21, 18 and 14 microg/mL; and bioassay 30, 15, 15 and 12 microg/mL (p = 0.1) . Both antibiotics eluted with a highly effective bactericidal activity . Our findings indicate that the presence of tobramycin has a synergistic-like effect on the bactericidal activity of vancomycin, which has not been previously reported . We recommend a combination of vancomycin and tobramycin with cement for the treatment of orthopedic infections caused by gram-positive organisms. Can J Vet Res, 2001 Jul, 65(3), 196 - 200 Experimental infection of specific pathogen free piglets with French strains of Streptococcus suis capsular type 2; Berthelot-Herault F et al.; A standardized model of Streptococcus suis type 2 infection in specific-pathogen-free piglets, housed in high-security barns, was used to compare the virulence of 3 French field strains of S . suis serotype 2 isolated from tonsils of a healthy pig (strain 65) or from diseased pigs (meningitis, strain 166', or septicemia, strain 24) . In one of the 2 trials, 7-week-old pigs, in 3 groups of 8, were inoculated intravenously with 2 x 10(8) colony-forming units of S . suis type 2 . In each group, 1 uninfected animal was a sentinel . Eight animals were also used as negative control group . The experiment was repeated under similar conditions with strains 65 and 166' . Virulence differed markedly among these S . suis strains when clinical signs, zootechnical performances, lesions, and bacteriological data were analyzed . Strain 65 did not induce clinical signs in inoculated pigs . In contrast, pigs infected with the other 2 strains exhibited clinical signs and typical lesions of S . suis type 2 infections . Differences in virulence were also observed between the 2 virulent strains . Sentinel animals exhibited the same manifestations as those recorded in inoculated piglets . Results were similar in the second trial, indicating that under the present experimental conditions, results were reproducible . The standardized conditions described in this study could be a useful tool to further study about the S . suis infection. Can J Vet Res, 2001 Jul, 65(3), 156 - 60 Decrease of the adhesion of Streptococcus suis serotype 2 mutants to embryonic bovine tracheal cells and porcine tracheal rings; Brassard J et al.; Streptococcus suis is an important swine pathogen that may be present in the tonsils of pigs that show no signs of illness . Because adhesion to host cells may be important in the carrier state, this study was undertaken to investigate adhesion to host cells by S . suis mutant strains defective in expression of a 39-kDa protein . Mutant strains of S . suis were generated by transposon Tn916 mutagenesis and were tested for adhesion to embryonic bovine tracheal cells and porcine tracheal rings . Compared with the parent strain, there was a significant reduction in adherence of 3 mutant strains to both bovine tracheal cells and porcine tracheal rings. Can J Vet Res, 2001 Jul, 65(3), 151 - 5 Prevalence, capsular type and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs in Korea; Han DU et al.; This study was undertaken to determine the prevalence, capsular serotype, and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs . Capsular serotype and antimicrobial susceptibility were determined by coagglutination test and agar dilution minimum inhibitory concentration, respectively . Streptococcus suis was isolated from 55 of the 406 palatine tonsillar samples tested (13.8%) and 14 of the 29 sampled herds (48.3%) . Of the 55 isolates recovered from slaughter pigs, 26 (47.3%) were untypeable . Of the remaining 29 isolates, capsular serotypes 9 (9 isolates) and 16 (4 isolates) were the most common, followed by capsular serotypes 4 (3 isolates) and 7 (3 isolates) . Every capsulated isolate was typeable and no palatine tonsillar sample yielded more than one serotype . Most of isolates were susceptible to low concentrations (MIC90) of amoxicillin (2 microg/mL), ceftiofur (1 microg/mL), and penicillin (1 microg/mL) . No correlation was found between antimicrobial susceptibility and capsular serotype. Hosp Med, 2001 Jul, 62(7), 406 - 9 Pneumococcal disease in childhood; Mohan PV et al.; Streptococcus pneumoniae is the most common cause of serious bacterial infections in children worldwide . Problems with antibiotic resistance have lead to changes in antibiotic policies for children with possible pneumococcal disease . Demonstration of the efficacy of a pneumococcal conjugate vaccine has lead to consideration of its inclusion in routine infant vaccination schedules. Hua Xi Kou Qiang Yi Xue Za Zhi, 1997 Feb, 15(1), 51 - 4 {An animal model for the mucosal immunization with the surface protein antigen P1 from streptococcus mutans and immune enhancement of cholera toxin}; Li F et al.; To study the immune enhancement of cholera toxin by developing an mucosal immune animal model, SD rats were immunized using different antigen composition through intragastric route (I.G) . The results showed that the effects of immunization became stronger after P1 was conjugated to cholera toxin B subunit (CTB) or procholeragenoid (PCG) . Even microgram dose of P1 could induce the rising of anti-P1 SIgA antibodies in the rat's saliva, but the antibodies risied little without this adjuvant . These implayed that the conjugation was much necessary in IG immunization . The immune enhancement of PCG was greater than that of CTB. Hua Xi Kou Qiang Yi Xue Za Zhi, 1997 Feb, 15(1), 48 - 50 {A primary observation of the active immune prevention effects against dental caries using the streptococcus mutans surface protein P1 in rats}; Li F et al.; To observe the anti-caries effects of a small dose of the surface protein antigen P1 . The SD rats were immunized with P1-CTB and P1-PCG through IN (intranasal immunization) and IG (intragastric immunization) . The results showed that caries was reduced heavily . There was no obvious difference between IN and IG . P1-CTB and P1-PCG can be used as an antigen to prevent the dental caries, but much further research work still need to do. Hua Xi Kou Qiang Yi Xue Za Zhi, 1997 Feb, 15(1), 45 - 7 {Conjugation and identification of the surface protein antigen P1 from streptococcus mutans MT6R with the cholera toxin B subunit and procholeragenoid}; Li F et al.; To make an effective antigen to prevent dental caries, the surface protein antigen P1(Ag I/II) was purified and identified by the rabbit anti-Ag I/II serum and was covalently conjugated to CTB and PCG by SPDP respectively . GM1-ELISA showed that both the conjugated P1-CTB and P1-PCG possess the ability to bind GM1 specifically without losing the antigenicity of P1 . This results showed that the conjugated antigens could be used in the immune prevention against dental caries. Hua Xi Kou Qiang Yi Xue Za Zhi, 1997 Nov, 15(4), 338 - 40 {Effects of different dilution of specific antibodies against the cell surface protein P1 of S . mutans MT 6R upon its adherence}; Zou J et al.; The sera and saliva were obtained from BALB/C mice by injection of the cell surface protein P1 and Freud's adjuvant . High antibody titers were showed in the sera and saliva by an enzyme-linked immunosorbent assay . Different dilution of the specific antibodies against the protein P1 of S . mutans had different inhibition effects on adhering of S . mutans MT 6R . Adherence of streptococcus mutans MT 6R on saliva-coated hydroxyapatite was reduced significantly by using sera and salivary antibodies in dilution of 1:1, 1:2, 1:4, 1:8, 1:16, 1:32 (P < 0.05). Hua Xi Kou Qiang Yi Xue Za Zhi, 1997 Nov, 15(4), 294 - 6, 311 {Study on the vaccine against dental caries of recombinant Streptococcus lactis and artificial active immunization . V . Extraction, purification and dot blot analysis of RNA in the recombinant S . lactis}; Ling J et al.; To investigate the transcription level of the pac gene in the recombinant S . lactis, the amount of PAc-specific mRNA was measured by RNA dot blot analysis with the biotin-labeled 1.5 kb PstI fragment of the pac gene . RNA were extracted with guanidinium thiocyanate followed by centrifugation in cesium chloride solutions . These results indicated the sequence of PAc-mRNA in S . lactis HL107 or HL45 was the same as that in S . mutans Ingbritt and showed similar density photograph . The PAc-specific mRNA plated an intermediary role of protein synthesis in recombinant S . lactis and made it complete the expression of cloned pac gene. Saudi Med J, 2001 Jul, 22(7), 565 - 8 Necrotizing fasciitis; Cherneski CL et al.; Necrotizing fasciitis is a devestating condition which has been recognized for several years . In North America a recent increase of cases has led to much media attention and public fear . Necrotizing fasciitis may occur as a consequence of infection with Streptococcus pyogenes or as a result of a polymicrobial synergistic infection caused by aerobic, anaerobic, gram positive and gram negative organisms, often in postoperative patients . Necrotizing fasciitis caused by Streptococcus pyogenes is mediated by superantigens . The management of necrotizing fasciitis requires a high index of suspicion for diagnosis followed by antimicrobial therapy and early surgical intervention . In cases caused by Streptococcus pyogenes with streptococcal toxic shock syndrome, intravenous immunoglobulin may be of benefit. FEMS Immunol Med Microbiol, 2001 Jul, 31(1), 21 - 7 Serum antibodies to group A streptococcal extracellular and cell-associated antigens in Egyptians with post-streptococcal diseases; Abdel-Rehim M et al.; We investigated serum antibodies to a comprehensive array of group A streptococcal antigens and superantigens in Egyptian subjects . Antibodies to Streptococcus pyogenes cell-associated proteins and to proteins released by rapidly dividing S . pyogenes were compared in four patient groups with different post-streptococcal diseases and in healthy controls . Enzyme-linked immunosorbent assays showed that total Ig and IgG to extracellular antigens were significantly higher in patients with acute rheumatic fever (ARF) compared to healthy controls, but no differences were found in either total Ig or IgG titres to cell-associated proteins between any of the groups . Western blotting showed that multiple extracellular and cell-associated antigens, covering a wide range of molecular masses, were recognised by all sera, including healthy controls . No evidence was obtained for putative dominant antigens associated with any disease group, although a low molecular mass cell-associated protein (approximately 4 kDa) was clearly recognised by two-thirds of subjects irrespective of disease status . These findings demonstrate that raised serum Ig and IgG titres to extracellular, but not cell-associated, S . pyogenes antigens are a feature of ARF in this population, and suggest that multiple S . pyogenes antigens contribute to this response. Medicina (B Aires), 2001, 61(3), 295 - 300 Phenotypic and genotypic study of Streptococcus agalactiae in vagina of pregnant women in Argentina; Toresani I et al.; Streptococcus agalactiae (GBS) is a leading cause of serious neonatal infection . In this study we determine the prevalence, serotype distribution and genomic diversity of GBS in vagina of pregnant women . Vaginal swabs of 531 pregnant women were cultured on Columbia Agar Base Blood, GBS Agar Base and Todd Hewitt Broth . GBS were characterized by group and type-specific agglutination . Genomic polymorphism was studied by random amplification of DNA (RAPD) . Seventeen patients (3.2%) were positive for GBS, resulting serotype III the most frequent . RAPD detected 16 different RAPD profiles from 21 GBS studied, revealing a good discriminatory power . In this sense, this method showed different genotype from GBS serotype III recovered from successive samples of two patients, suggesting reinfection . In conclusion, the combination of RAPD and serotyping appear promising for epidemiological studies . Finally, findings of reinfection after therapy during pregnancy, led us to suggest performing prenatal GBS screening and intrapartum prophylaxis in order to reduce neonatal risk. J Antimicrob Chemother, 2001 Jul, 48(1), 67 - 74 Efficacy and safety of gemifloxacin in the treatment of community-acquired pneumonia: a randomized, double-blind comparison with trovafloxacin; File TM Jr et al.; This multicentre, randomized, double blind, parallel group study compared the efficacy and safety of gemifloxacin (320 mg once daily) with trovafloxacin (200 mg once daily) in 571 patients with community-acquired pneumonia (CAP) . Although treatment was given routinely for 7 days it could be extended to 14 days; two-thirds of patients were treated for 7 days . High clinical success rates were noted at follow-up in the per-protocol population in both the gemifloxacin group (95.8%) and the trovafloxacin group (93.6%), non-inferiority with 95% CI . In the intent-to-treat population, the clinical success rate at follow-up was significantly superior for gemifloxacin (87.6%) compared with trovafloxacin (81.1%; 95% CI 0.5, 12.4) . The pathogens identified most commonly at presentation were Mycoplasma pneumoniae and Streptococcus pneumoniae . Gemifloxacin eradicated 100% of S . pneumoniae . One bacteraemic isolate of S . pneumoniae was associated with clinical failure in the trovafloxacin group (MIC of trovafloxacin 8 mg/L) . Gemifloxacin was well tolerated and the incidence of transient liver function abnormalities was very low . Gemifloxacin is an effective and well-tolerated treatment for patients with CAP. J Infect Dis, 2001 Sep 1, 184(5), 591 - 6 Epub 2001 Jul 24. National Department of Defense Surveillance for Invasive Streptococcus pneumoniae: antibiotic resistance, serotype distribution, and arbitrarily primed polymerase chain reaction analyses; Hudspeth MK et al.; To provide surveillance among US military personnel and their beneficiaries, 157 invasive Streptococcus pneumoniae clinical isolates were collected systematically from 7 large military hospitals between August 1997 and August 1999 . The isolates were studied for antibiotic resistance, and 120 were serotyped and subjected to arbitrarily primed polymerase chain reaction (AP-PCR) . Fifty (31.9%) of 157 isolates had intermediate or high-level resistance to penicillin, and 15.9% had multidrug resistance . The most common serotypes were 4, 6B, 9V, 14, 19F, and 23F . Those serotypes associated with penicillin resistance were 6B, 9V, 19A, and 19F . Most invasive disease cases were caused by serotypes included in the currently available 23- and 7-valent pneumococcal vaccines . By use of AP-PCR, 4 DNA groups were correlated with health care site (P< or =.0001) . These results are valuable in assessing appropriate use of antibiotics and vaccines against S . pneumoniae in both military personnel and their families. J Clin Microbiol, 2001 Aug, 39(8), 3006 - 8 Rapid pulsed-field gel electrophoresis method for group B streptococcus isolates; Benson JA et al.; We developed a rapid pulsed-field gel electrophoresis (PFGE) method that required 3 days to complete, an improvement over the standard method that required as many as 8 days . The accuracy and reproducibility of the rapid method were verified by analysis of DNA band sizes of our control group B streptococcus isolate . The rapid method was superior to the standard method, providing more precise molecular sizing and gels of higher image quality . The reproducibility of rapid PFGE substantiated its value and continued use. Appl Environ Microbiol, 2001 Aug, 67(8), 3756 - 8 Recovery of Streptococcus iniae from diseased fish previously vaccinated with a streptococcus vaccine; Bachrach G et al.; Streptococcus iniae was recovered from diseased rainbow trout (Oncorhynchus mykiss, Walbaum) previously vaccinated against streptococcosis . PCR and serological methods indicate the presence of a new serotype in the diseased fish. Appl Environ Microbiol, 2001 Aug, 67(8), 3396 - 405 Altered protein expression of Streptococcus oralis cultured at low pH revealed by two-dimensional gel electrophoresis; Wilkins JC et al.; Streptococcus oralis is the predominant aciduric nonmutans streptococcus isolated from the human dentition, but the role of this organism in the initiation and progression of dental caries has yet to be established . To identify proteins that are differentially expressed by S . oralis growing under conditions of low pH, soluble cellular proteins extracted from bacteria grown in batch culture at pH 5.2 or 7.0 were analyzed by two-dimensional (2-D) gel electrophoresis . Thirty-nine proteins had altered expression at low pH; these were excised, digested with trypsin using an in-gel protocol, and further analyzed by peptide mass fingerprinting using matrix-assisted laser desorption ionization mass spectrometry . The resulting fingerprints were compared with the genomic database for Streptococcus pneumoniae, an organism that is phylogenetically closely related to S . oralis, and putative functions for the majority of these proteins were determined on the basis of functional homology . Twenty-eight proteins were up-regulated following growth at pH 5.2; these included enzymes of the glycolytic pathway (glyceraldehyde-3-phosphate dehydrogenase and lactate dehydrogenase), the polypeptide chains comprising ATP synthase, and proteins that are considered to play a role in the general stress response of bacteria, including the 60-kDa chaperone, Hsp33, and superoxide dismutase, and three distinct ABC transporters . These data identify, for the first time, gene products that may be important in the survival and proliferation of nonmutans aciduric S . oralis under conditions of low pH that are likely to be encountered by this organism in vivo. Pediatr Int, 2001 Aug, 43(4), 385 - 90 Investigation of risk factors for penicillin-resistant Streptococcus pneumoniae carriage in Turkish children; Ciftci E et al.; BACKGROUND: Nasopharyngeal colonization plays an important role for infections caused by Streptococcus pneumoniae . Emergence of penicillin resistance in this organism has made it difficult to treat pneumococcal infections . The objectives of this study were to investigate the risk factors for nasopharyngeal colonization with S . pneumonia and for nasopharyngeal colonization with penicillin-resistant S . pneumoniae . METHODS: Three hundred children with or without evidence of infection were investigated for various risk factors . Streptococcus pneumoniae isolated from children's nasopharyngeal swabs were examined for penicillin susceptibility . RESULTS: Day-care attendance (odds ratio OR=2.82, P=0.003) and upper respiratory tract infection within the last month (OR=1.83, P=0.02), have been determined to be risk factors for S . pneumoniae carriage . The use of antibiotics within the last 3 months (OR=81.07, P<0.001), the presence of more than five people living in the house of the child (OR=6.63, P=0.03), and having a sibling under 5-years-old (OR=4.60, P=0.03) have been determined to be risk factors for penicillin-resistant S . pneumoniae carriage . CONCLUSION: Some children are inevitably exposed to and colonized with penicillin susceptible or resistant S . pneumoniae . Changes in day-care organizations, better living conditions, and restriction of antibiotic use seems to be useful precautions to prevent the emerging and colonization with penicillin-susceptible or -resistant S . pneumoniae. Lett Appl Microbiol, 2001 Aug, 33(2), 122 - 5 Evaluation of co-aggregation among Streptococcus mitis, Fusobacterium nucleatum and Porphyromonas gingivalis; Nagayama M et al.; AIMS: To develop a semi-quantitative method for evaluating co-aggregation reactions among three bacterial species, and to examine the influence of Fusobacterium nucleatum on the adherence of Porphyromonas gingivalis . METHODS AND RESULTS: The method involves coating hydroxyapatite (HAP) discs with streptococcal cells and treatment with radio-labelled bacterial cell suspensions . The sensitivity of the method was estimated by comparison with a turbidometric co-aggregation assay . Results from the two methods were in close agreement . Streptococcus mitis-coated HAP discs were immersed in a 3H-labelled Fus . nucleatum cell suspension and then a 14C-labelled P . gingivalis cell suspension . The discs were then pyrolysed to recover and quantify the released 3H and 14C radioactivity . The number of Fus . nucleatum cells on the discs increased with immersion time and this, in turn, resulted in elevated adherence of P . gingivalis . CONCLUSION: The data indicate that the method closely reflects co-aggregation characters, and that Fus . nucleatum has a positive effect on the adherence of P . gingivalis . SIGNIFICANCE AND IMPACT OF THE STUDY: The present method, which is designed to mimic the oral environment, should prove useful in the semi-quantitative evaluation of co-aggregation reactions. J Vet Med B Infect Dis Vet Public Health, 2001 Jun, 48(5), 347 - 55 Epidemiological relationship of human and swine Streptococcus suis isolates; Tarradas C et al.; Two cases of meningitis due to Streptococcus suis in humans are reported here . A butcher and an abattoir worker were referred to a health centre in Castellon (Spain) with fever and symptoms of meningitis . After adequate treatment, a slight hipoacusia persisted as sequelae in both cases . Colonies of S . suis group R, serotype 2 and phenotype MRP+EF+ were isolated from cerebroespinal fluid . Epidemiological studies showed that both workers had in common the handling of pork meat of slaughtered healthy pigs from three closed farms . A study of the tonsils from apparently healthy, slaughtered pigs was carried out . A total of 234 tonsillar samples were obtained and 81 strains of S . suis were isolated from them . Serotype 2 appeared to be the most frequent (50.6%), and the analysis for phenotype showed a high percentage of tonsillar strains with the phenotype MRP+EF+ (35.9%) . The humans and 28 tonsillar swine strains showed a similar profile (S . suis group R, serotype 2 and phenotype MRP+EF+) . A total of 26 of the swine isolates were analysed by ribotyping using EcoRI . The human strains showed the same six-band hybridization pattern that shared five bands with the pattern most frequently shown by most of the tonsillar N . suis group R, serotype 2 and phenotype MRP+EF+ strains, differing only in the lightest, faintest band which was slightly less anodical in human (> or = 1.8 kb) than in swine (approximately 1.8 kb) . From these results, both groups of strains, humans and porcine, showed differences; how can these differences in the pattern of ribotyping be explained if they should have the same origin? Is it possible that they have undergone an adaptation to the new host or perhaps the modification is due to other unknown causes? Further studies in this area are required in order to answer these questions. Biosci Biotechnol Biochem, 2001 Jun, 65(6), 1290 - 5 Purification and characterization of an oligo-isomaltosaccharide synthase from a Streptococcus sobrinus glucosyltransferase-I deficient mutant; Shinozaki-Kuwahara N et al.; One glucosyltransferase (GTF) -I deficient mutant of Streptococcus sobrinus strain B13N was isolated through chemical mutagenesis with ethyl methanesulfonate, and characterized . This mutant, designated as B13N-Id, readily allowed us to purify a homogeneous oligo-isomaltosaccharide synthase (GTF-S) from its culture fluid . The purified GTF-S was only recognized with rabbit polyclonal antibody against recombinant GTF-S from an Ecsherichia coli MD124 clone expressing the B13N gtfS gene, and showed the almost same enzymatic properties as the recombinant enzyme . A double reciprocal plot of the B13N GTF-S for sucrose was biphasic, and the affinity for this substrate was high compared to that of GTF-S enzymes from other strains. Ann Trop Paediatr, 2001 Jun, 21(2), 175 - 8 Group A beta-haemolytic streptococcal acute chest event in a child with sickle cell anaemia; Suara RO; Acute chest syndrome is a major cause of death and hospitalisation in children with sickle cell anaemia . It is often initiated by an infection, particularly pneumonia . Microbial agents previously not associated with acute chest syndrome are becoming increasingly important . Group A beta-haemolytic Streptococcus (GABHS) is thought to be an uncommon cause of pneumonia in children with sickle cell anaemia . We report a 15-year-old African-American girl who presented with an acute chest event characterised by fever, cough, chest pain, shortness of breath, right upper abdominal quadrant pain, jaundice and otitis media . Chest radiograph showed multi-lobar pneumonia with left pleural effusion . Group A beta-haemolytic Streptococcus was isolated from culture of pleural and middle ear fluids . She responded to therapy that included antibiotics, exchange blood transfusion, oxygen, thoracotomy chest tube drainage and decortication . In a child with sickle cell anaemia presenting with fever and an acute chest event, pneumonia should be considered and GABHS recognised as a possible aetiological agent . In addition, a chest X-ray should be obtained and antibiotics against agents causing community-acquired pneumonia instituted. Ann Trop Paediatr, 2001 Jun, 21(2), 149 - 54 Antimicrobial resistance of nasopharyngeal isolates of Streptococcus pneumoniae in healthy carriers: report of a study in 5-year-olds in Marcory, Abidjan, Côte d'Ivoire; Kacou-N'Douba A et al.; The increasing prevalence of antibiotic-resistant Streptococcus pneumoniae is of growing public health concern . The aim of this study was to assess resistance rates of S . pneumoniae to penicillin and other antimicrobial agents . Between November 1997 and February 1998 in a community health centre in Marcory, an Abidjan suburb, 138 S . pneumoniae strains were isolated from the nasopharynxes of 218 apparently healthy children aged 3-60 months . The sensitivity of the isolates was tested using the Kirby-Bauer method . In isolates with a possibly abnormal sensitivity to the Kirby-Bauer test, minimum inhibitory concentrations (MIC) were estimated using the E-test . Antimicrobials tested included penicillin G, amoxycillin, cefotaxime, cotrimoxazole, tetracycline, chloramphenicol, erythromycin, rifampicin and vancomycin . Twelve of 108 isolates (8.7%) had reduced sensitivity to penicillin G, and in three of them the MIC for penicillin reached at least 2 micrograms/ml . Resistance to amoxycillin and cefotaxime was lower than to penicillin (2.2%) . With regard to cotrimoxazole, 37% were moderately resistant and 15.2% highly resistant . The lowest resistance rate observed was to rifampicin (2.2%) and the highest was to tetracycline (57.2%) . Rates of resistance to erythromycin and chloramphenicol were 11.6% and 2.9% . All strains were sensitive to vancomycin . Multidrug resistance (MDR) was detected in 9.4% of S . pneumoniae isolates . In children, epidemiological surveillance of resistance can be monitored by bacteriological surveys, as shown in this study. J Infect Dis, 2001 Aug 15, 184(4), 451 - 9 Epub 2001 Jul 06. Nasopharyngeal carriage of Streptococcus pneumoniae in Finnish children younger than 2 years old; Syrjanen RK et al.; To describe the natural course of nasopharyngeal carriage of Streptococcus pneumoniae and its relationship to acute otitis media (AOM), 329 Finnish children were followed from ages 2 to 24 months . In total, 3024 nasopharyngeal (NP) swabs (obtained at 10 scheduled healthy visits) and 2007 NP aspirates (obtained during respiratory infections) were cultured . Carriage during health increased gradually (9%-43%) with age . Within 4 age intervals, carriage was lower during health (13%-43%) than during respiratory infection without AOM (22%-45%) . Higher proportions of positive samples were found during AOM (45%-56%), in particular during pneumococcal AOM (97%-100%) . Antimicrobial treatment reduced carriage only temporarily . The most frequent NP serotypes were 6B, 6A, 11, 19F, and 23F . Both age and health status were important determinants of NP carriage of S . pneumoniae and these features should be considered carefully during analysis of carriage rates. J Biol Chem, 2001 Sep 21, 276(38), 35606 - 13 Epub 2001 Jul 23. Specific interactions between F1 adhesin of Streptococcus pyogenes and N-terminal modules of fibronectin; Ensenberger MG et al.; Protein F1 is a surface protein of Streptococcus pyogenes that mediates high affinity binding to fibronectin (Fn) and facilitates S . pyogenes adherence and penetration into cells . The smallest portion of F1 known to retain the full binding potential of the intact protein is a stretch of 49 amino acids known as the functional upstream domain (FUD) . Synthetic and recombinant versions of FUD were labeled with fluorescein isothiocyanate and used in fluorescence anisotropy experiments . These probes bound to Fn or the 70-kDa fragment of Fn with dissociation constants of 8-30 nm . Removal of the N-terminal seven residues of FUD did not cause a change in binding affinity . Further N- or C-terminal truncations resulted in complete loss of binding activity . Analysis of recombinant versions of the 70-kDa fragment that lacked one or several type I modules indicates that residues 1-7 of the 49-mer bind to type I modules I1 and I2 of the 27-kDa subfragment and the C-terminal residues bind to modules I4 and I5 . Fluorescein isothiocyanate-labeled 49-mer also bound with lower affinity to large Fn fragments that lack the five type I modules of the 27-kDa fragment but contain the other seven type 1 modules of Fn . These results indicate that, although FUD has a general affinity for type I modules, high affinity binding of FUD to Fn is mediated by specific interactions with N-terminal type I modules. Int J Infect Dis, 2001, 5(2), 86 - 8 Comparative analysis of outcome and clinical features in children with pleural empyema caused by penicillin-nonsusceptible and penicillin-susceptible Streptococcus pneumoniae; Paganini H et al.; OBJECTIVE: To describe the clinical characteristics of pleural empyema in children caused by Streptococcus pneumoniae nonsusceptible to penicillin and compare their clinical outcome with pleural empyemas caused by penicillin-susceptible organisms . METHODS: Records of 109 children with complicated pleural effusions between January 1996 and December 1998 were retrospectively reviewed . RESULTS: Pathogens were recovered in 45 of these cases . Streptococcus pneumoniae represented 71% of the isolates, with 20 (62%) and 12 (38%) of the strains proving susceptible and nonsusceptible to penicillin, respectively . The average age for children with penicillin-nonsusceptible infections was 18.3 months compared with an average age of 40.9 months for those with penicillin-susceptible infections (P = 0.03) . Previous antibiotic treatment was more frequent in children with penicillin-nonsusceptible organisms (P = 0.05) . No significant differences were found between the two groups for gender, underlying diseases, duration of fever and tachypnea, need of surgical treatment, bacteremia incidence, mean duration of therapy, or length of hospital stay . CONCLUSIONS: Pleural empyemas caused by penicillin-nonsusceptible S . pneumoniae appear to be associated with younger age and previous antibiotic treatment . By contrast, the outcome was not influenced by the penicillin susceptibility of the pneumococcus strains. J Bacteriol, 2001 Aug, 183(16), 4927 - 31 The licC gene of Streptococcus pneumoniae encodes a CTP:phosphocholine cytidylyltransferase; Rock CO et al.; The licC gene product of Streptococcus pneumoniae was expressed and characterized . LicC is a nucleoside triphosphate transferase family member and possesses CTP:phosphocholine cytidylyltransferase activity . Phosphoethanolamine is a poor substrate . The LicC protein plays a role in the biosynthesis of the phosphocholine-derivatized cell wall constituents that are critical for cell separation and pathogenesis. Pediatr Infect Dis J, 2001 Jul, 20(7), 711 - 4 Nocardiosis in South Texas children; Fergie JE et al.; Primary cutaneous nocardiosis can present as cellulitis or abscess with or without lymphadenitis . In South Texas children with suspected Staphylococcus aureus or Streptococcus pyogenes skin and soft tissue infections who do not improve as expected with appropriate antibiotics, physicians should consider Nocardia brasiliensis in the differential diagnosis, especially if there are multiple lesions in a lymphocutaneous distribution. Science, 2001 Jul 20, 293(5529), 498 - 506 Complete genome sequence of a virulent isolate of Streptococcus pneumoniae; Tettelin H et al.; The 2,160,837-base pair genome sequence of an isolate of Streptococcus pneumoniae, a Gram-positive pathogen that causes pneumonia, bacteremia, meningitis, and otitis media, contains 2236 predicted coding regions; of these, 1440 (64%) were assigned a biological role . Approximately 5% of the genome is composed of insertion sequences that may contribute to genome rearrangements through uptake of foreign DNA . Extracellular enzyme systems for the metabolism of polysaccharides and hexosamines provide a substantial source of carbon and nitrogen for S . pneumoniae and also damage host tissues and facilitate colonization . A motif identified within the signal peptide of proteins is potentially involved in targeting these proteins to the cell surface of low-guanine/cytosine (GC) Gram-positive species . Several surface-exposed proteins that may serve as potential vaccine candidates were identified . Comparative genome hybridization with DNA arrays revealed strain differences in S . pneumoniae that could contribute to differences in virulence and antigenicity. Int J Antimicrob Agents, 2001 Jul, 18(1), 9 - 17 Bacteriological and clinical efficacy of various antibiotics used in the treatment of streptococcal pharyngitis in Italy . An epidemiological study; Rondini G et al.; A total of 123 community paediatricians and 23 microbiology laboratories studied the clinical and bacteriological efficacy of treatment of group A streptococcal pharyngitis in Italy . Of 1065 patients, from whom Streptococcus pyogenes was isolated, 723 returned to follow up and of these 138 (19%) still had a positive throat culture . The erythromycin resistance (ER) rate was 23.7% with resistance phenotype distribution of: 31.7% constitutive (CR), 26.6% inducible (IR) and 41.7% efflux pump (M) resistance phenotype . All strains were susceptible to the beta-lactam agents tested . CR strains were highly resistant to all 14, 15 and 16 membered macrolides with the exception of rokitamycin which showed activity against 37.8% of isolates . All phenotype M and some IR isolates were susceptible to clindamycin, rokitamycin, josamycin and spiramycin; clarithromycin was active against a small percentage of strains belonging to the IR and M phenotype . Bacterial eradication was found in 85.5, 78.7 and 75.8% of the penicillin, macrolide and cephalosporin treated groups . Genotyping of strains showed that 8.7% of the 19% of cases classified as 'failed bacterial eradication' were due to recolonization with a different isolate, observed exclusively among beta-lactams treated patients . Clinical cure was achieved in a high percentage of cases, irrespective of the antibiotic prescribed, with the best clinical efficacy being found following therapy with amoxycillin and clarithromycin (90.9%). Clin Infect Dis, 2001 Aug 15, 33(4), 517 - 22 Epub 2001 Jul 20. A nosocomial outbreak of fluoroquinolone-resistant Streptococcus pneumoniae; Weiss K et al.; Over the course of a 20-month period, in a hospital respiratory ward in which ciprofloxacin was often used as empirical antimicrobial therapy for lower respiratory tract infections (LRTIs), 16 patients with chronic bronchitis developed nosocomial LRTIs caused by penicillin- and ciprofloxacin-resistant Streptococcus pneumoniae (serotype 23 F) . The minimum inhibitory concentration (MIC) of ciprofloxacin for all isolates from the first 9 patients was 4 microg/mL, in association with a parC mutation . Isolates from the subsequent 7 patients all had a ciprofloxacin MIC of 16 microg/mL, in association with an additional mutation in gyrA . The MICs for this isolate were 8 microg/mL of levofloxacin (resistant), 2 microg/mL of moxifloxacin and gatifloxacin (intermediately resistant), and 0.12 microg/mL of gemifloxacin . This outbreak demonstrates the ability of S . pneumoniae to acquire multiple mutations that result in increasing levels of resistance to the fluoroquinolones and to be transmitted from person to person. Clin Infect Dis, 2001 Aug 15, 33(4), 483 - 8 Epub 2001 Jul 20. Macrolide-resistant Streptococcus pneumoniae and use of antimicrobial agents; Pihlajamaki M et al.; The prevalence of isolates of Streptococcus pneumoniae (pneumococcus) that are resistant to antimicrobial agents is increasing globally . We studied the connection between antimicrobial resistance of pneumococci and regional use of antimicrobial agents in Finland . In 1997, a total of 6106 pneumococcal isolates were identified in clinical microbiology laboratories in Finland . Most of the pneumococci were isolated from respiratory tract samples, 8% were from blood culture samples, and 0.5% were from cerebrospinal fluid samples . The regional levels of resistance for pneumococci in 1997 were compared with the regional rates of use of antimicrobial agents from 1995 through 1996 . We found that resistance to macrolides correlated highly significantly with macrolide use (P=.006) . A significant correlation was also found between resistance to trimethoprim-sulfamethoxazole and trimethoprim-sulfamethoxazole use (P=.043) . We did not find a correlation between penicillin resistance and the use of any antimicrobial agent . The positive correlation between macrolide-resistant pneumococci and the use of macrolides is worrying, because macrolides are used worldwide in the treatment of patients with respiratory tract infections, which are often caused by pneumococci. Mol Genet Metab, 2001 Jul, 73(3), 259 - 67 Impairment of type III group B Streptococcus-stimulated superoxide production and opsonophagocytosis by neutrophils in diabetes; Mazade MA et al.; The effect of hyperglycemia upon susceptibility to bacterial infection in diabetes mellitus is incompletely elucidated . The present experiments assessed the effect of hyperglycemia upon neutrophil-mediated phagocytosis of type III group B Streptococcus (GBS) . Type III GBS was chosen for study because the incidence of invasive GBS disease is substantially increased in type 2 diabetic compared with nondiabetic subjects . The hypothesis tested was that severe hyperglycemia would alter neutrophil metabolism by diverting NADPH from superoxide production into the aldose reductase-dependent polyol pathway that converts glucose into sorbitol and thus would impair opsonophagocytosis (OP) of type III GBS . Neutrophils from 10 adults with type 2 diabetes had no intrinsic phagocytic defect under baseline glycemic conditions . After equilibration in 60 or 120 mM glucose or in 60 mM choline chloride, OP activity was reduced significantly (P < or = 0.03) . Neutrophil superoxide production correlated with glucose concentration and also was significantly reduced during hyperglycemia (P < 0.05) . Addition of III GBS capsular polysaccharide-specific IgG in a sufficient concentration supported efficient OP, even during hyperglycemia . Alrestatin, an aldose reductase inhibitor, increased superoxide production and significantly improved OP of type III GBS (P = 0.03) . Thus, diversion of NADPH into the polyol pathway is one mechanism by which OP of GBS III is impaired during hyperglycemia, and this effect is mitigated when levels of capsular polysaccharide-specific IgG are sufficient . J Clin Dent, 2000, 11(1), 16 - 9 An evaluation of resin-ionomers to prevent coronal microleakage in endodontically treated teeth; Nup C et al.; While a gutta-percha system is most commonly used by practitioners for canal obturation, there is no agreement on how best to close the chamber of an endodontically treated tooth . Some materials used in restorative dentistry may have endodontic applications . Restorative materials of particular interest to endodontists are those used as subgingival restorations . Their properties include insolubility in oral fluids, adhesiveness, dual-cure capabilities, radiopacity, hardness and fluoride release, low cure shrinkage and a low coefficient of thermal expansion . The purpose of this in vitro study was to test three products that could be used to seal pulp chambers of endodontically treated teeth to prevent bacterial infiltration . A bacteriological assay system was used to determine the efficacy of three commercially available restorative materials to prevent penetration of Streptococcus salivarius from the pulp chambers into the prepared canals . The materials evaluated were: Geristore two-paste system with Tenure Quik with fluoride, Dispersalloy with Tenure Quik with fluoride, and Marathon posterior composite with Tenure Quik with fluoride . During the sixty days of the study, the analysis indicated that the Geristore two-paste system with Tenure Quik with fluoride provided a statistically significant improved seal when compared to the Marathon posterior composite and Dispersalloy amalgam with Tenure Quik with fluoride. Mol Cells, 2001 Jun 30, 11(3), 360 - 8 Molecular cloning and characterization of groESL operon in Streptococcus pneumoniae; Kim SN et al.; GroEL is a major target of the immune defense in infection and seems to be negatively regulated by HrcA in gram-positive organisms . However, HrcA's mechanism has not been elucidated . To elucidate the role of groEL in Streptococcus pneumoniae, the groESL operon was cloned in Escherichia coli . The promoter region of the pneumococcal groESL operon contained a sigmaA type promoter and an inverted repeat (CIRCE) . A Northern blot analysis of the groESL operon demonstrated that the groESL operon is transcribed as a bicistronic mRNA, and reached maximum expression 7.5 to 10 min after heat shock . A primer extension analysis showed a potential transcription start point at 155 bp upstream of the translation start site, preceding the groES gene . The putative negative regulator of the groEL gene, hrcA, of S . pneumoniae was recovered by PCR-based chromosomal walking from grpE locus . A sequence analysis showed a sigmaA type promoter flanked by 2 CIRCE elements . His-tagged HrcA was overexpressed as a soluble form in E . coli and bound to the CIRCE regions in the promoter of both groESL and dnaK operons in vitro . Additionally, a helix-loop helix motif, a putative DNA binding domain, was found at the C-terminal of HrcA . These results will help to determine the nature of HrcA in the groESL repression. BMC Microbiol . 2001;1(1):9 . Epub 2001 Jul 02. Sub-inhibitory concentrations of vancomycin prevent quinolone-resistance in a penicillin-resistant isolate of Streptococcus pneumoniae; Cottagnoud P et al.; BACKGROUND: The continuous spread of penicillin-resistant pneumococci represents a permanent threat in the treatment of pneumococcal infections, especially when strains show additional resistance to quinolones . The main objective of this study was to determine a treatment modality impeding the emergence of quinolone resistance . RESULTS: Exposure of a penicillin-resistant pneumococcus to increasing concentrations of trovafloxacin or ciprofloxacin selected for mutants resistant to these drugs . In the presence of sub-inhibitory concentrations of vancomycin, development of trovafloxacin-resistance and high-level ciprofloxacin-resistance were prevented . CONCLUSIONS: Considering the risk of quinolone-resistance in pneumococci, the observation might be of clinical importance. Int J Dermatol, 2001 Apr, 40(4), 285 - 7 Psoriasis vulgaris and acute guttate psoriasis in a family; Banno T et al.; A 33-year-old man (Case 1) developed scaling and erythema on the extensor side of his extremities, head, and back, 3 years before his first visit to our hospital . The lesions spread out to form several plaques and became persistent (Fig . 1a) . Serum antistreptolysin O (ASO) was within the normal range and a pharyngeal culture showed normal flora . Histologic findings were compatible with psoriasis vulgaris (Fig . 1b) . Topical steroid therapy was effective; however, scaling and erythema persisted . The 8-year-old daughter (Case 2) of Case 1 had suffered severe tonsillitis 2 months prior to the skin eruptions . Numerous rain drop-like scaly papules appeared on her legs and generalized rapidly (Fig . 2a) . Her tonsils were swollen and reddish . Streptococcus pyogenes was detected by pharyngeal smear . The ASO value was at a high level . Histologic examination showed slightly elongated rete ridges, focal lack of the granular layer, parakeratosis, mild spongiosis, and exostosis of neutrophils (Fig . 2b) . She was diagnosed with acute guttate psoriasis . Administration of oral antimicrobials reduced the ASO value and led to remission of the disease . After 1 month of antimicrobial treatment, the erythematous papules disappeared . The human leukocyte antigen (HLA) typing of the father was HLA-A24, A31, B7, B39, and Cw7, and that of the daughter was HLA-A24, A31, B39, and Cw7 . Both possessed HLA-A24, B39, and Cw7 . The HLA typing of other family members was also studied . The mother had HLA-A11, A24, B39, B62, Cw4, and Cw7, and her son had HLA-A11, A24, B7, B62, Cw4, and Cw7 . The result of HLA typing is shown in Table 1 . Lymphocyte stimulating test was performed using heat-killed S . pyogenes which were boiled at 70 degrees C for 10 min and sonicated . The stimulation index was twofold higher in the daughter than in the father (Fig . 3). Microb Pathog, 2001 Aug, 31(2), 81 - 9 Identification of a major, CsrRS-regulated secreted protein of Group A streptococcus; Heath A et al.; CsrR/CsrS (CovR/CovS) is a two-component regulator of extracellular virulence factors in Group A streptococcus, but the full range of regulated exoproteins is unknown . Since CsrR represses expression of regulated factors, culture supernates of wild-type and CsrR(-)mutant strains were compared by two-dimensional gel electrophoresis (2DGE) to identify regulated exoproteins . Supernates of DeltacsrRS(-)mutant, but not wild-type, bacteria contained an abundant 23 kDa protein . The N-terminal sequence of this spot corresponded to a putative open reading frame (ORF) in the streptococcal genome . In a mobility shift assay, phosphorylated CsrR bound to a PCR amplicon that included sequences upstream of this ORF . By primer extension analysis, the ORF (designated mspA, for Mucoidy-associated Secreted Protein) was expressed in mid- and late-exponential phase in a DeltacsrRS(-)mutant . The presence of an in-frame deletion in mspA did not affect colony appearance, mucoidy or in vitro growth, and there was no difference between DeltamspA and wild-type strains in a mouse model of skin infection . MspA is co-regulated with other factors required for dermonecrosis (e.g . capsule, streptolysin S and purogenic exotoxin B); however, deletion of this gene does not affect expression of hyaluronic acid capsule or severity of skin infection in mice . Antimicrob Agents Chemother, 2001 Aug, 45(8), 2400 - 2 Antibiotic susceptibility and mechanisms of erythromycin resistance in clinical isolates of Streptococcus agalactiae: French multicenter study; De Mouy D et al.; Among 126 Streptococcus agalactiae isolates collected in 10 French laboratories in 1999, 27 (21.4%) had macrolide resistance related to the presence of erm(B) (11 strains), erm(A) subclass erm(TR) (10 strains), and mef(A) genes (2 strains) and the presence of combinations of erm(B) and erm(A) genes or mef(A) genes (3 strains). Antimicrob Agents Chemother, 2001 Aug, 45(8), 2393 - 6 Alterations in MurM, a cell wall muropeptide branching enzyme, increase high-level penicillin and cephalosporin resistance in Streptococcus pneumoniae; Smith AM et al.; We report that alteration in MurM, an enzyme involved in the biosynthesis of branched-stem cell wall muropeptides, is required for maximal expression of penicillin and cefotaxime resistance in the pneumococcus . Hungarian isolate 3191 (penicillin MIC, 16 microg/ml; cefotaxime MIC, 4 microg/ml) was a source of donor DNA in transformation experiments . Penicillin-binding protein DNA was insufficient to transform recipient strain R6 to full resistance . Further transformation with altered murM DNA was required for full expression of donor penicillin and cefotaxime resistance. Antimicrob Agents Chemother, 2001 Aug, 45(8), 2375 - 7 Prevalence of macrolide resistance genes in clinical isolates of the Streptococcus anginosus ("S . milleri") group; Jacobs JA et al.; Twenty-two unrelated erythromycin-resistant anginosus group strains (3.2% resistance rate) were assessed for mechanisms of resistance . Streptococcus anginosus accounted for 16 of the 22 isolates . Fifteen isolates harbored the erm(B) gene . The erm(TR) and the mef(E) genes were carried by two isolates each . In three isolates, none of these resistance genes was detected by PCR. Antimicrob Agents Chemother, 2001 Aug, 45(8), 2309 - 15 Kappacin, a novel antibacterial peptide from bovine milk; Malkoski M et al.; Caseinomacropeptide (CMP) is a heterogeneous C-terminal fragment (residues 106 to 169) of bovine milk kappa-casein composed of glycosylated and phosphorylated forms of different genetic variants . We have demonstrated that CMP has growth-inhibitory activity against the oral opportunistic pathogens Streptococcus mutans and Porphyromonas gingivalis and against Escherichia coli . CMP was fractionated using reversed-phase high-performance liquid chromatography (RP-HPLC), and each fraction was tested for activity against S . mutans in a 96-well-plate broth assay . Fractions were characterized by N-terminal sequence analysis and mass spectrometry . The active form of CMP was shown to be the nonglycosylated, phosphorylated kappa-casein (residues 106 to 169) {kappa-casein(106--169)}, which we have designated kappacin . Endoproteinase Glu-C was used to hydrolyze CMP, and the generated peptides were separated using RP-HPLC and gel filtration-HPLC and then tested for activity against S . mutans . The peptide Ser(P)(149)kappa-casein-A(138--158) was the only peptide generated by endoproteinase Glu-C digestion that exhibited growth-inhibitory activity . Peptides corresponding to the sequences of the inhibitory peptide Ser(P)(149)kappa-casein-A(138--158) and its nonphosphorylated counterpart kappa-casein-A(138--158) were chemically synthesized and tested for antibacterial activity . The synthetic Ser(P)(149) kappa-casein-A(138--158) displayed growth-inhibitory activity against S . mutans (MIC, 59 microg/ml {26 microM}) . The nonphosphorylated peptide, however, did not inhibit growth at the concentrations tested, indicating that phosphorylation is essential for activity. Indian J Pediatr, 2001 Jun, 68(6), 561 - 2 Mixed anaerobic and aerobic testicular abscess in a neonate; Singh D et al.; A premature baby developed a testicular abscess on day 28 of life . The pus aspirated from the abscess grew a mixture of beta hemolytic Streptococcus and a Bacteroides species . The source of this infection could not be identified . The baby improved on antibiotic therapy . This is the first reported case of a polymicrobial testicular infection involving an anaerobe in a neonate . The relevant literature is reviewed. Arch Pediatr Adolesc Med, 2001 Jun, 155(6), 687 - 91 Performance of a predictive model for streptococcal pharyngitis in children; Attia MW et al.; CONTEXT: Group A beta-hemolytic streptococcus (GABHS) pharyngitis is a common childhood illness . The clinical diagnosis is difficult to determine and laboratory tests have limitations; hence, the condition is generally overdiagnosed and overtreated . Several clinical pediatric-specific predictive models have been published but none have been prospectively studied . OBJECTIVE: To test the performance of a previously published predictive model for GABHS pharyngitis in children in different clinical settings and during different seasons . DESIGN: Prospective cohort study . SETTINGS: Pediatric emergency department and 2 pediatric outpatient clinics . PATIENTS: Children aged between 1 and 18 years with pharyngitis on initial examination at study sites between April 1, 1999, and March 31, 2000 . INTERVENTIONS: Recording of clinical features during initial evaluation using a standardized form and recovery of GABHS from patients' throats using reference standard methods . MAIN OUTCOME MEASURES: Posttest probability for GABHS positive throat culture associated with the model's positive predictors (moderate to severe tonsillar swelling, cervical lymphadenopathy {moderate to severe tenderness and enlargement of cervical lymph nodes}, scarletiniform rash, and the absence of coryza) and the models' negative predictors (absence of the above signs and the presence of coryza) . RESULTS: Of 587 patients analyzed, 218 (37%) had a positive throat culture for GABHS . Forty-nine percent were boys . Mean +/- SD age was 6.7 +/- 3.9 years . There was no difference between the subsets within the sample . The posttest probability values for a positive throat culture associated with positive and negative predictors of the model were 79% and 12%, respectively . CONCLUSIONS: A pediatric predictive model for GABHS pharyngitis performed better than physicians' subjective estimates for a positive throat culture and was comparable with a rapid antigen detection test . The model performed consistently well in different populations and across seasons . It can be useful if reliable microbiological testing and/or follow-up are not attainable. Eur J Immunol, 2001 Jul, 31(7), 2104 - 15 The protein tyrosine kinase inhibitor AG126 prevents the massive microglial cytokine induction by pneumococcal cell walls; Hanisch UK et al.; Central nervous system (CNS) infections caused by Streptococcus pneumoniae still have a disastrous outcome . Underlying immunological and CNS cellular events are largely enigmatic . We used pneumococcal cells walls (PCW) to investigate microglial responses as these cells are prominent sensors and effectors during neuropathological changes . PCW stimulation of mouse microglia in vitro evoked the release of the cyto- and chemokines, TNF-alpha, IL-6, IL-12, KC, MCP-1, MIP-1alpha, MIP-2 and RANTES as well as soluble TNF receptor II, a potential TNF-alpha antagonist . The release induction followed extremely steep dose-response relations, and short exposure periods (15 min) were already sufficient to trigger substantial responses . PCW signaling controlling the release depended on both p38 and p42/p44 (ERK2/ERK1) MAP kinase activities . The kinase inhibitor, tyrphostin AG126 prevented the PCW-inducible phosphorylation of p42/p44(MAPK), potently blocked cytokine release and drastically reduced the bioavailable TNF-alpha, since it only marginally affected the release of soluble TNF receptors . Moreover, in an in vivo model of pneumococcal meningitis, AG126 significantly attenuated the PCW-induced leukocyte influx to the cerebrospinal fluid . The findings imply that pneumococcal CNS infection can cause a rapid and massive microglial activation and that ERK/MAPK pathway(s) are potential targets for pharmacological interventions. Infect Immun, 2001 Aug, 69(8), 5162 - 5 Localization of surface immunogenic protein on group B streptococcus; Rioux S et al.; The localization and accessibility of the group B streptococcus (GBS) surface immunogenic protein (Sip) at the surface of intact GBS cells were studied by flow cytometric assay and immunogold electron microscopy . Antibodies present in pooled sera collected from mice after immunization with purified recombinant Sip efficiently recognized native Sip at the surfaces of the different GBS strains tested, which included representatives of all nine serotypes . Examination of GBS cells by immunogold electron microscopy revealed that the Sip-specific antibodies attached preferentially to polar sites and the septal region . This result confirmed that Sip is exposed at the intact-cell surface, but it also suggests that its distribution is restricted to certain regions of the cell. Infect Immun, 2001 Aug, 69(8), 4767 - 73 Facilitated intranasal induction of mucosal and systemic immunity to mutans streptococcal glucosyltransferase peptide vaccines; Smith DJ et al.; Synthetic peptide vaccines which are derived from functional domains of Streptococcus mutans glucosyltransferases (GTF) have been shown to induce protective immunity in Sprague-Dawley rats after subcutaneous injection in the salivary gland region . Since mucosal induction of salivary immunity would be preferable in humans, we explored methods to induce mucosal antibody in the rat to the GTF peptide vaccines HDS and HDS-GLU after intranasal administration . Several methods of facilitation of the immune response were studied: the incorporation of peptides in bioadhesive poly(D,L-lactide-coglycolide) (PLGA) microparticles, the use of monoepitopic (HDS) or diepitopic (HDS-GLU) peptide constructs, or the use of mucosal adjuvants . Salivary immunoglobulin A (IgA) responses were not detected after intranasal administration of diepitopic HDS-GLU peptide constructs in alum or after incorporation into PLGA microparticles . However, significant primary and secondary salivary IgA and serum IgG antibody responses to HDS were induced in all rats when cholera holotoxin (CT) or a detoxified mutant Escherichia coli heat-labile enterotoxin (R192G LT) were intranasally administered with HDS peptide constructs in PLGA . Coadministration of LT with HDS resulted in predominantly IgG2a responses in the serum, while coadministration with CT resulted in significant IgG1 and IgG2a responses to HDS . Serum IgG antibody, which was induced to the HDS peptide construct by coadministration with these adjuvants, also bound intact mutans streptococcal GTF in an enzyme-linked immunosorbent assay and inhibited its enzymatic activity . Thus, immune responses which are potentially protective for dental caries can be induced to peptide-based GTF vaccines after mucosal administration if combined with the CT or LT R192G mucosal adjuvant. Glycobiology, 2001 Jul, 11(7), 587 - 92 Cell surface-expressed Thomsen-Friedenreich antigen in colon cancer is predominantly carried on high molecular weight splice variants of CD44; Singh R et al.; Increased mucosal expression of TF, the Thomsen-Friedenreich oncofetal blood group antigen (galactose beta1-3 N-acetylgalactosamine alpha-) occurs in colon cancer and colitis . This allows binding of TF-specific lectins, such as peanut agglutinin (PNA), which is mitogenic to the colorectal epithelium . To identify the cell surface TF-expressing glycoprotein(s), HT29 and Caco2 colon cancer cells were surface-labeled with Na{(125)I} and subjected to PNA-agarose affinity purification and electrophoresis . Proteins, approximately 110-180 kDa, present in HT29 but not Caco2 were identified by Western blotting as high molecular weight splice variants of CD44 (CD44v) . Selective removal of TF antigen by Streptococcus pneumoniae endo-alpha-N-acetylgalactosaminidase substantially reduced PNA binding to CD44v . Immunoprecipitated CD44v from HT29 cell extracts also expressed sialyl-Tn (sialyl 2-6 N-acetylgalactosaminealpha-) . Incubation of PNA 15 microg/ml with HT29 cells caused no additional proliferative effect in the presence of anti-CD44v6 mAb . In colon cancer tissue extracts (N = 3) PNA bound to CD44v but not to standard CD44 . These data show that CD44v is a major PNA-binding glycoprotein in colon cancer cells . Because CD44 high molecular weight splice variants are present in colon cancer and inflammatory bowel disease tissue but are absent from normal mucosa, these results may also explain the increased PNA reactivity in colon cancer and inflammatory bowel disease . The coexpression of oncofetal carbohydrate antigens TF and sialyl-Tn on CD44 splice variants provides a link between cancer-associated changes in glycosylation and CD44 splicing, both of which correlate with increased metastatic potential. J Pediatr, 2001 Jul, 139(1), 148 - 51 A mutation in Bruton's tyrosine kinase as a cause of selective anti-polysaccharide antibody deficiency; Wood PM et al.; Children and adults can have recurrent infection with invasive encapsulated bacterial pathogens such as Streptococcus pneumoniae as a result of a selective inability to respond to polysaccharide antigens . We have identified a mutation in the gene encoding Bruton's tyrosine kinase in a male patient with selective anti-polysaccharide antibody deficiency. FEBS Lett, 2001 Jul 6, 500(3), 141 - 4 A novel approach for purification of recombinant proteins using the dextran-binding domain; Kaseda K et al.; Using the dextran-binding domain (DBD) of a type of glucosyltransferase (GTF) from Streptococcus sobrinus, we have developed a novel method for purifying recombinant proteins . DBD-tagged green and red fluorescent proteins as well as the parent GTF and DBD moiety were adsorbed well to commercially available cross-linked dextran (such as Sephadex beads and Sephacryl beads), and eluted efficiently with water-soluble dextran . The purity of the eluted proteins after this one-step affinity purification was approximately 90% or better . The results suggest that DBD can be used as a powerful carrier for purification of various recombinant proteins. J Matern Fetal Med, 2001 Jun, 10(3), 203 - 8 Evaluation of a rapid optical immunoassay-based test for group B streptococcus colonization in intrapartum patients; Samadi R et al.; OBJECTIVE: To compare an optical immunoassay (OIA) rapid diagnostic kit to standard culture for the diagnosis of vaginal colonization with group B streptococcus (GBS) and to assess the accuracy and reproducibility of the OIA results . METHOD: A total of 301 patients in labor were prospectively evaluated for GBS colonization with a test approved by the Food and Drug Administration (STREP B OIA kit, Biostar, Boulder, CO, USA) and by culture . The vagina was simultaneously sampled with two swabs . Rectal culture was obtained separately . RESULTS: By the criterion of a positive culture, the vagina was colonized by GBS in 33 of 301 (11%) patients; and the rectum in 42 of 301 (13.9%) . The vagina or rectum or both were colonized by CBS in 54 of 301 (17.9%) of patients . The OIA had sensitivity, specificity, positive predictive value and negative predictive value of 63.6%, 86.3%, 37.5% and 94.8%, respectively . The OIA had a kappa statistic score of 0.59 . CONCLUSION: The OIA is not an adequately sensitive rapid kit for reliable detection of GBS colonization of the vagina . The results of the OIA were only moderately accurate and reproducible. J Infect Dis, 2001 Aug 1, 184(3), 292 - 300 Epub 2001 Jun 26. Microbiological and inflammatory factors associated with the development of pneumococcal pneumonia; Dallaire F et al.; Pneumococcal pneumonia still is associated with a high mortality rate, despite appropriate antimicrobial therapy . Many gaps remain in the understanding of the pathogenesis of this deadly infection . The microbial and inflammatory events that characterize survival or death after intranasal inoculation of mice with an LD(50) inoculum of Streptococcus pneumoniae were investigated . Survival was associated with rapid bacterial clearance and low inflammation (surfactant and red blood cells in alveoli), but no neutrophil recruitment or lung tissue injury was noted . By contrast, death was preceded by strong bacterial growth that peaked 48 h after the infection and was associated with gradual increases in pulmonary levels of interleukin-6, macrophage inflammatory protein (MIP)-1alpha, MIP-2, monocyte chemoattractant protein-1, KC, and neutrophil recruitment . The injection of tumor necrosis factor-alpha or the addition of lipopolysaccharide or heat-killed S . pneumoniae to the inoculum enhanced early host response and survival . These observations may help develop appropriate markers of evolution of pneumonia, as well as new therapeutic strategies. J Infect Dis, 2001 Aug 1, 184(3), 285 - 91 Epub 2001 Jun 26. Antibodies to capsular polysaccharides of group B Streptococcus in pregnant Canadian women: relationship to colonization status and infection in the neonate; Davies HD et al.; In a cohort study of 1207 pregnant women in Alberta, Canada, the serotype distributions of vaginal-rectal group B Streptococcus (GBS) isolates were compared with all isolates from neonates with invasive GBS disease identified by population-based surveillance . Serum concentrations of Ia, Ib, II, III, and V capsular polysaccharide (CPS)-specific IgG also were determined, according to serotype of the vaginal-rectal colonizing GBS strain . GBS colonization was detected in 19.5% (235 of 1207) of women . Serotype III accounted for 20.6% (48 of 233) of colonizing strains available for typing but for 37% (27 of 73) of invasive isolates from neonates (P<.01) . Maternal colonization with type III was least likely to be associated with moderate concentrations of III CPS-specific IgG . Serotype III GBS is more invasive than other serotypes in this population; this may be due, at least in part, to poor maternal type III CPS-specific antibody response. J Infect Dis, 2001 Aug 1, 184(3), 268 - 77 Epub 2001 Jun 26. The etiology of community-acquired pneumonia at an urban public hospital: influence of human immunodeficiency virus infection and initial severity of illness; Park DR et al.; In a prospective study, the etiology of community-acquired pneumonia (CAP) was investigated among consecutive patients admitted to an academic, urban public hospital in Seattle . The study population was uniquely young, was predominantly male, and had high rates of homelessness, cigarette smoking, alcoholism, injection drug use, and human immunodeficiency virus (HIV) infection . Leading causes of CAP among HIV-negative patients were aspiration, followed by Streptococcus pneumoniae, Legionella species, and Mycoplasma pneumoniae . Among HIV-positive patients, Pneumocystis carinii, Mycobacterium tuberculosis, S . pneumoniae, and M . pneumoniae were the most common etiologic agents . Severe CAP was associated with typical bacterial infections and aspiration pneumonia but not Legionella infection among HIV-negative patients and with Pseudomonas aeruginosa infections among HIV-positive patients . These findings emphasize the need to tailor empirical antibiotic therapy according to local patient populations and individual risk factors and highlight the importance of recognizing underlying HIV infection in patients who are hospitalized with CAP. J Bacteriol, 2001 Aug, 183(15), 4599 - 608 Competence repression under oxygen limitation through the two-component MicAB signal-transducing system in Streptococcus pneumoniae and involvement of the PAS domain of MicB; Echenique JR et al.; In Streptococcus pneumoniae, a fermentative aerotolerant and catalase-deficient human pathogen, oxidases with molecular oxygen as substrate are important for virulence and for competence . The signal-transducing two-component systems CiaRH and ComDE mediate the response to oxygen, culminating in competence . In this work we show that the two-component MicAB system, whose MicB kinase carries a PAS domain, is also involved in competence repression under oxygen limitation . Autophosphorylation of recombinant MicB and phosphotransfer to recombinant MicA have been demonstrated . Mutational analysis and in vitro assays showed that the C-terminal part of the protein and residue L100 in the N-terminal cap of its PAS domain are both crucial for autokinase activity in vitro . Although no insertion mutation in micA was obtained, expression of the mutated allele micA59DA did not change bacterial growth and overcame competence repression under microaerobiosis . This was related to a strong instability of MicA59DA-PO(4) in vitro . Thus, mutations which either reduced the stability of MicA-PO(4) or abolished kinase activity in MicB were related to competence derepression under microaerobiosis, suggesting that MicA-PO(4) is involved in competence repression when oxygen becomes limiting . The micAB genes are flanked by mutY and orfC . MutY is an adenine glycosylase involved in the repair of oxidized pyrimidines . OrfC shows the features of a metal binding protein . We did not obtain insertion mutation in orfC, suggesting its requirement for growth . It is proposed that MicAB, with its PAS motif, may belong to a set of functions important in the protection of the cell against oxidative stress, including the control of competence. Mol Microbiol, 2001 Jun, 40(6), 1427 - 38 Unique regulation of SclB - a novel collagen-like surface protein of Streptococcus pyogenes; Rasmussen M et al.; Slipped-strand mispairing at sites containing so-called coding repeats (CRs) can lead to phase variation of surface proteins in Gram-negative bacteria . This mechanism, believed to contribute to virulence, has so far not been identified in a Gram-positive bacterium . In the genome of the Gram-positive human pathogen Streptococcus pyogenes, we identified pentanucleotide CRs within a putative signal sequence of an open reading frame (ORF) encoding a novel collagen-like surface protein, denoted SclB . In 12 S . pyogenes strains, the number of CRs in the sclB gene varied from three to 19, rendering the start codon in frame with the downstream ORF in four strains and out of frame in eight strains . A protein reacting with anti-SclB antibodies could only be solubilized from three strains, all containing an intact sclB gene . Variations in the number of CRs were observed within strains of the same M serotype and occurred during growth of S . pyogenes in fresh human blood, but not in medium . The SclB protein has a hypervariable N-terminal part, a collagen-like central part and a typical cell wall sorting sequence containing the LPXTGX motif . SclB is related to the collagen-like SclA and is, like SclA, involved in the adhesion of S . pyogenes bacteria to human cells . However, the Mga protein, known to upregulate sclA and several additional genes encoding virulence factors of S . pyogenes, downregulates sclB transcription . This observation and the potential of SclB to phase vary by slipped-strand mispairing emphasize the unique regulation of this novel S . pyogenes surface protein. Mol Microbiol, 2001 Jun, 40(6), 1273 - 87 alpha-Enolase of Streptococcus pneumoniae is a plasmin(ogen)-binding protein displayed on the bacterial cell surface; Bergmann S et al.; Binding of human plasminogen to Streptococcus pneumoniae and its subsequent activation promotes penetration of bacteria through reconstituted basement membranes . In this study, we have characterized a novel pneumococcal surface protein with a molecular mass of 47 kDa, designated Eno, which specifically binds human plasmin(ogen), exhibits alpha-enolase activity and is necessary for viability . Using enzyme assays, we have confirmed the alpha-enolase activity of both pneumococcal surface-displayed Eno and purified recombinant Eno protein . Immunoelectron microscopy indicated the presence of Eno in the cytoplasm as well as on the surface of encapsulated and unencapsulated pneumococci . Plasminogen-binding activity was demonstrated with whole pneumococcal cells and purified Eno protein . Binding of activated plasminogen was also shown for Eno; however, the affinity for plasmin is significantly reduced compared with plasminogen . Results from competitive inhibition assays indicate that binding is mediated through the lysine binding sites in plasmin(ogen) . Carboxypeptidase B treatment and amino acid substitutions of the C-terminal lysyl residues of Eno indicated that the C-terminal lysine is pivotal for plasmin(ogen)-binding activity . Eno is ubiquitously distributed among pneumococcal serotypes, and binding experiments suggested the reassociation of secreted Eno to the bacterial cell surface . The reassociation was also confirmed by immunoelectron microscopy . The results suggest a mechanism of plasminogen activation for human pathogens that might contribute to their virulence potential in invasive infectious processes. Mol Microbiol, 2001 Jun, 40(6), 1249 - 59 Sites of pH regulation of the urea channel of Helicobacter pylori; Weeks DL et al.; Helicobacter pylori (Hp) and Streptococcus salivarius (Ss) require intrabacterial urease for acid resistance and express a urea channel, UreI . The presence of UreI was shown to increase urea permeability approximately 300-fold over that of a non-polar ureI deletion mutant . Expression of SsUreI in Xenopus oocytes increased urea uptake pH independently, whereas HpUreI shows an acidic pH dependence, half-maximal at pH 6.0 . Mutagenesis of all histidines, aspartates, glutamates and the lysine in the periplasmic domain of HpUreI showed that His-123, His-131, Asp-129, Asp-140, Glu-138 and Lys-132 in the second periplasmic loop (PL2) and His-193 in the C-terminus (Ct) were important for activation of transport . With the exception of a lysine that was shown to substitute for His-193 in HpUreI, these charged amino acids are absent in SsUreI . A chimera in which PL1 of HpUreI was replaced by PL1 of SsUreI retained activity at acidic pH and gained partial activity at neutral pH . Exchange of PL2 inactivated transport, whereas exchange of Ct had no effect . Chimeras, in which either PL1 or PL2 of HpUreI replaced those of SsUreI, retained wild-type transport, but replacement of the Ct or both loops inactivated transport . PL1 appears to be important for restricting transport through HpUreI at neutral pH, whereas protonation of three histidines in PL2 and Ct and the presence of three dicarboxylic amino acids in PL2 appears to be necessary to activate HpUreI at acidic pH. J Vet Pharmacol Ther, 2001 Apr, 24(2), 83 - 8 Depletion study of trimethoprim and sulphadiazine in milk and its relationship with mastitis pathogenic bacteria strains minimum inhibitory concentrations (MICs) in dairy cows; Nunez BS et al.; Time-related concentrations in milk of a combination of trimethoprim-sulphadiazine (TMP-SDZ) intramammary formulated infusion and its relationship with pathogenic bacteria strains minimum inhibitory concentrations (MICs) isolated from clinical mastitis cows were analysed . The MICs study was performed for Escherichia coli, Staphylococcus aureus and Streptococcus sp . strains . The SDZ concentrations in milk were analysed using high-performance liquid chromatography (HPLC) and TMP using a microbiological assay . Ten lactating cows milked three times daily were used in the time-concentration studies of TMP-SDZ . Milk samples (approximately 20 mL) from the treated mammary quarters were taken at 6, 12, 24, 30 and 36 h after first administration . In order to define the withdrawal time, milk samples from the treated mammary quarters were taken at 24, 36, 48, 72, 84 and 96 h, after finishing the therapy . The MICs fluctuated between 1 and 8 microg/mL . Effective therapeutic concentrations lasted for 36 h when intramammary infusion was repeated three times every 12 h . No TMP was detected in milk for 24 h after finishing therapy . Milk SDZ concentrations were below 0.1 microg/mL in all treated cows after 84 h finishing therapy . At 96 h after finishing therapy, no SDZ milk concentrations were found in six animals, although four animals of the experimental group still had concentrations of 0.07 microg/mL. J Invest Dermatol, 2001 Jul, 117(1), 91 - 7 Cutaneous injury induces the release of cathelicidin anti-microbial peptides active against group A Streptococcus; Dorschner RA et al.; Cathelicidins are a family of peptides thought to provide an innate defensive barrier against a variety of potential microbial pathogens . The human and mouse cathelicidins (LL-37 and CRAMP, respectively) are expressed at select epithelial interfaces where they have been proposed to kill a number of gram-negative and gram-positive bacteria . To determine if these peptides play a part in the protection of skin against wound infections, the anti-microbial activity of LL-37 and CRAMP was determined against the common wound pathogen group A Streptococcus, and their expression was examined after cutaneous injury . We observed a large increase in the expression of cathelicidins in human and murine skin after sterile incision, or in mouse following infection by group A Streptococcus . The appearance of cathelicidins in skin was due to both synthesis within epidermal keratinocytes and deposition from granulocyctes that migrate to the site of injury . Synthesis and deposition in the wound was accompanied by processing from the inactive prostorage form to the mature C-terminal peptide . Analysis of anti-microbial activity of this C-terminal peptide against group A Streptococcus revealed that both LL-37 and CRAMP potently inhibited bacterial growth . Action against group A Streptococcus occurred in conditions that typically abolish the activity of anti-microbial peptides against other organisms . Thus, cathelicidins are well suited to provide defense against infections due to group A Streptococcus, and represent an important element of cutaneous innate immunity. Microb Drug Resist, 2001 Summer, 7(2), 99 - 125 Annotated draft genomic sequence from a Streptococcus pneumoniae type 19F clinical isolate; Dopazo J et al.; The public availability of numerous microbial genomes is enabling the analysis of bacterial biology in great detail and with an unprecedented, organism-wide and taxon-wide, broad scope . Streptococcus pneumoniae is one of the most important bacterial pathogens throughout the world . We present here sequences and functional annotations for 2.1-Mbp of pneumococcal DNA, covering more than 90% of the total estimated size of the genome . The sequenced strain is a clinical isolate resistant to macrolides and tetracycline . It carries a type 19F capsular locus, but multilocus sequence typing for several conserved genetic loci suggests that the strain sequenced belongs to a pneumococcal lineage that most often expresses a serotype 15 capsular polysaccharide . A total of 2,046 putative open reading frames (ORFs) longer than 100 amino acids were identified (average of 1,009 bp per ORF), including all described two-component systems and aminoacyl tRNA synthetases . Comparisons to other complete, or nearly complete, bacterial genomes were made and are presented in a graphical form for all the predicted proteins. Microb Drug Resist, 2001 Summer, 7(2), 165 - 70 Macrolide resistance in Streptococcus pyogenes isolates from throat infections in the region of Aachen, Germany; Brandt CM et al.; Macrolide-resistance was assessed in 216 consecutive Streptococcus pyogenes isolates from throat infections in the region of Aachen, Germany . Seventeen isolates were resistant to erythromycin: 12 isolates revealed a macrolide (M) phenotype and harbored mefA, and five strains expressed an inducible macrolide-lincosamide-streptogramin B (MLSB) phenotype of which four strains harbored ermA(TR) and one strain contained ermB(AM) . Telithromycin (HMR 3647) and quinupristin/dalfopristin remained active particularly against the ermA(TR)-containing S . pyogenes isolates studied . Random amplified polymorphic DNA analysis identified multiple clones among erythromycin-resistant strains, but did not discriminate beyond the emm-type . mefA was present in three isolates either with emm2, emm12, or emm75, and in nine isolates with emm4 . All four strains with ermA(TR) contained emm77, and the single strain with ermB(AM) harbored emm1 . Despite the relative low rate of macrolide-resistance, these data suggest that at least three different macrolide-resistance determinants are prevalent in Germany and that mefA has spread rapidly into multiple clones of S . pyogenes. Microb Drug Resist, 2001 Summer, 7(2), 153 - 64 Presence of international multiresistant clones of Streptococcus pneumoniae in Colombia; Vela MC et al.; In Colombia, penicillin resistance of Streptococcus pneumoniae invasive isolates recovered from children less than 5 years old has increased from 10% in 1994 to 49.4% in 1999, suggesting the circulation of international resistant clones in the country . A total of 167 S . pneumoniae invasive isolates with diminished susceptibility to penicillin (DSP) were studied . The techniques used were pulsed-field gel electrophoresis (PFGE) and restriction fragment length polymorphism (RFLP) of penicillin-binding proteins (PBPs) genes (2B, 2X, and 1A) . Forty-nine serotype 23F isolates were grouped in two clusters: 15 (31%) multiresistant isolates showed PFGE pattern A and PBP I profile, thus making them indistinguishable from Spain23F-1 clone, and 34 (69%) with PFGE pattern C, PBP II profile, and intermediate level resistance (ILR) to penicillin and TMP-SMX, features unique to a Colombian clone . Fifty-five serotype 14 isolates were assigned to PFGE B pattern, PBP III profile, having high-level resistance to penicillin, and TMP-SMX, similar to the France9V variant 14 . This same pattern was present in five capsular type 9V isolates . Four serotype 14 isolates were assigned to PFGE pattern F, and appeared to be similar to Slovakia(14)-10 PFGE pattern, although they had different PBP profiles . Nine capsular type 6B and one 6A isolates belonged to PFGE pattern M, similar to Spain6B-2, although they showed different PBP profiles . The remaining 44 isolates, corresponding to serotypes 14, 6B, 19F, and 34, showed variable PFGE and PBP patterns . These results show that as many as two international clones may be circulating in Colombia as well as a unique, widely distributed 23F clone with ILR to penicillin . Additionally, some Colombian isolates capsular type 14 and 6B might be related to Slovakia(14)-10 Spain6B-2 clones, respectively. Microb Drug Resist, 2001 Summer, 7(2), 137 - 52 Penicillin-resistant Streptococcus pneumoniae in metropolitan New York hospitals: case control study and molecular typing of resistant isolates; Roberts RB et al.; During the 4-month period from January to April, 1998, 476 patients with Streptococcus pneumoniae infections were detected in 12 metropolitan New York hospitals and 112 penicillin-resistant (PRP) isolates (24%) were identified in 11 institutions . A case control study of 100 patients with penicillin-resistant and susceptible pneumococci from four of the widely dispersed hospitals revealed a high incidence of underlying medical illnesses in adult patients (74%), a preponderance of patients with pneumonia (63%), and a majority of patients who had underlying risk factors for pneumonia or invasive disease (51%) . In this limited case control study, no difference was noted between cases and controls regarding known risk factors for penicillin-resistant pneumococcal infections . The percentage of single-patient PRP isolates varied among individual hospitals but the mean percentages of PRP from the four participating University Medical Centers and seven community hospitals were similar: 26% and 22% respectively . By E-test, 60% and 26% were high-level penicillin and ceftriaxone resistant, respectively . Pulsed-field gel electrophoresis identified 26 chromosomal macrorestriction patterns among the 103 PRP isolates available for analysis, but almost half (50 isolates or 48%) of these belong to two drug-resistant internationally spread clones, SP(23)-1 and SP(9/14)-3, that were detected in all hospitals and were recovered from invasive and noninvasive sites in both children and adults. Ther Umsch, 2001 Jun, 58(6), 367 - 71 {Travelers' diarrhea}; Markwalder K; Diarrhea is the most common health problem of travelers to tropical destinations, affecting up to over 50%, with however considerable regional and seasonal variation . Orally acquired bacterial pathogens, particularly enterotoxigenic Escherichia coli, are the most frequent etiology of travelers' diarrhea occurring during the first three weeks of travel . Protozoal infections, e.g . giardia and Entamoeba histolytica, are more often the cause of diarrhea and prolonged problems of intestinal motility of the returning traveler--as are postinfectious irritable bowel syndromes . Prevention seems theoretically simple by avoiding any potentially contaminated food and drinks, but the principle of 'cook it, boil it, peel it, or avoid it is obviously a goal difficult to achieve . Several antibiotics have shown to be able to prevent diarrhea for a short period of time, but the potential of adverse effects and selection of resistant pathogens calls for a restrictive use for short trips of particularly vulnerable subjects only . The use of probiotics--e.g . Saccharomyces boulardi, Streptococcus faecium--gave conflicting results--both in prevention and treatment . The basics of treatment is appropriate fluid replacement--mostly by the oral route . Although this measure can safely bridge the time until spontaneous remission, it fails to reduce the duration of illness . Appropriate antibiotics are fairly effective to reduce the duration of travelers' diarrhea, especially if combined with loperamid . The administration of the later is contraindicated in small children . The most commonly used and well documented antibiotics belong to the fluoroquinolones . Alternatives for pediatric use are azithromycin and cotrimoxazole . Considering the mostly short duration of travelers' diarrhea the administration of antibiotics can be limited to cases of acute febrile dysentery and violent diarrhea when rapid relief is essential . In cases of febrile diarrhea malaria must be considered if the patient has been exposed to the risk of transmission. J Immunol, 2001 Jul 15, 167(2), 691 - 8 A genetic model of stress displays decreased lymphocytes and impaired antibody responses without altered susceptibility to Streptococcus pneumoniae; Murray SE et al.; Stress pathways affect immune function, the most notable of these pathways being activation of the hypothalamic-pituitary-adrenal (HPA) axis . Although HPA activation has generally been relegated to an immunosuppressive role, recent evidence suggests that stress and HPA activation can be immunoenhancing in certain situations . To investigate specific effects of stress on immune function, we used a genetic model of chronic stress wherein transgenic mice overexpress corticotropin-releasing hormone (CRH), a primary mediator of the stress response . In these mice, CRH is overproduced in the brain, leading to chronic activation of the HPA axis . We found that CRH-transgenic mice have decreased leukocyte numbers in lymphoid compartments, with preferential loss of B lymphocytes . They also exhibit decreased Ab production and impaired isotype switching in response to immunization with a thymus-dependent Ag, phosphocholine-keyhole limpet hemocyanin . Despite these deficits, immunization protected CRH-transgenic and wild-type mice equally well against lethal challenge with Streptococcus pneumoniae, an encapsulated Gram-positive bacterium known to require Ab-mediated opsonization for clearance . While IgG responses are severely depressed in these mice, IgM titers are only modestly decreased . This fairly robust IgM response may be sufficient to protect against S . pneumoniae . Additionally, while total leukocyte numbers are decreased in these mice, neutrophil numbers are increased . This increase in number of neutrophils may compensate for the depressed IgG response, allowing adequate host defense during chronic stress. Infection, 2001 May-Jun, 29(3), 163 - 5 Typing of Streptococcus pyogenes strains isolated from throat infections in the region of Aachen, Germany; Brandt CM et al.; BACKGROUND: Changes in the epidemiology of Streptococcus pyogenes infections may be associated with the introduction and reappearance of individual serotypes within a population . MATERIALS AND METHODS: Typing of 216 consecutive isolates of S . pyogenes from patients with pharyngitis in the region of Aachen, Germany, was performed by sequencing the emm gene, slide-agglutination of the T-antigen and determining the serum opacity reaction (SOR) . RESULTS: All 216 isolates were unequivocally emm-typable . emm1 was most common (18.5%), foLlowed by emm12 (15.7%), emm3 (14.4%) and emm28 (13.9%) . Only four isolates contained newly validated emm types: emm89 or emm94 were harbored by two isolates each . In one isolate, the sequence type s104 was found . CONCLUSION: Despite an anticipated selective pressure, the prevalence of emm1 among isolates from throat infections in northwestern Germany remains high, but does not reflect the predominance of emm1 among invasive isolates in Germany. Scand J Infect Dis, 2001, 33(5), 393 - 4 Capsular polysaccharide types of Streptococcus pneumoniae isolated from maxillary sinus effusion; Tahkokallio O et al.; Pneumococcal capsular polysaccharide vaccines may be a novel way of treating patients with recurrent sinusitis . To evaluate the pneumococcal capsular polysaccharide types of strains causing sinusitis, 55 pneumococcal strains isolated from maxillary sinus effusion were analyzed . The most frequently occurring types were 3 (13%), 19F (11%), 6B (9%) and 23F (9%). Scand J Infect Dis, 2001, 33(5), 381 - 3 Polymicrobial polyarticular septic arthritis: a rare clinical entity; Gilad J et al.; Polymicrobial polyarticular septic arthritis is a rare clinical entity, with only a few cases having been reported to date . We report a case due to Streptococcus pyogenes and Staphylococcus aureus in an IVDU, complicated by fatal streptococcal toxic-shock syndrome, and review the current literature . We conclude that whenever polymicrobial polyarticular septic arthritis is diagnosed, a high index of suspicion should be maintained for the detection of locally destructive infectious processes as well as systemic complications, and that a high mortality rate should be expected. Bull World Health Organ, 2001, 79(6), 528 - 33 Group A streptococcal sore throat in a periurban population of northern India: a one-year prospective study; Nandi S et al.; OBJECTIVE: To estimate the incidence and risk factors of group A streptococcus (GAS) sore throat among school-aged children living in a periurban slum area of Chandigarh, North India . METHODS: A total of 536 children aged 5-15 years from 261 families identified by a systematic random selection method were enrolled in the study . Episodes of sore throat were recorded through fortnightly home visits over a one-year period . The local vernacular (Hindi) terms gala kharab (bad throat) and khansi jukam (cough and cold) were used to identify symptoms of sore throat, and throat swab specimens were collected from children who had these symptoms on the day of the home visit . Bacterial culture was carried out and the isolation of GAS was confirmed using group-A-specific antiserum . FINDINGS: The incidences of sore throat and GAS sore throat were, respectively, 7.05 and 0.95 episodes per child-year . The incidence was higher in the following situations: among 11-year-olds, during the winter (November to January) and rainy (August) months (a bimodal peak), among children living in houses where there was no separate room for the kitchen, and in homes that included a tobacco smoker . CONCLUSION: The results show that the incidence of GAS sore throat was related to age, season, and indoor air pollution. JAMA, 2001 Jul 4, 286(1), 49 - 56 Effect of short-course, high-dose amoxicillin therapy on resistant pneumococcal carriage: a randomized trial; Schrag SJ et al.; CONTEXT: Emerging drug resistance threatens the effectiveness of existing therapies for pneumococcal infections . Modifying the dose and duration of antibiotic therapy may limit the spread of resistant pneumococci . OBJECTIVE: To determine whether short-course, high-dose amoxicillin therapy reduces risk of posttreatment resistant pneumococcal carriage among children with respiratory tract infections . DESIGN AND SETTING: Randomized trial conducted in an outpatient clinic in Santo Domingo, Dominican Republic, October 1999 through July 2000 . PARTICIPANTS: Children aged 6 to 59 months who were receiving antibiotic prescriptions for respiratory tract illness (n = 795) . INTERVENTIONS: Children were randomly assigned to receive 1 of 2 twice-daily regimens of amoxicillin: 90 mg/kg per day for 5 days (n = 398) or 40 mg/kg per day for 10 days (n = 397) . MAIN OUTCOME MEASURES: Penicillin-nonsusceptible Streptococcus pneumoniae carriage, assessed in nasopharyngeal specimens collected at days 0, 5, 10, and 28; baseline risk factors for nonsusceptible pneumococcal carriage; and adherence to regimen, compared between the 2 groups . RESULTS: At the day 28 visit, risk of penicillin-nonsusceptible pneumococcal carriage was significantly lower in the short-course, high-dose group (24%) compared with the standard-course group (32%); relative risk (RR), 0.77; 95% confidence interval (CI), 0.60-0.97; P =.03; risk of trimethoprim-sulfamethoxazole nonsusceptibility was also lower in the short-course, high-dose group (RR, 0.77; 95% CI, 0.58-1.03; P =.08) . The protective effect of short-course, high-dose therapy was stronger in households with 3 or more children (RR, 0.72; 95% CI, 0.52-0.98) . Adherence to treatment was higher in the short-course, high-dose group (82% vs 74%; P =.02) . CONCLUSION: Short-course, high-dose outpatient antibiotic therapy appears promising as an intervention to minimize the impact of antibiotic use on the spread of drug-resistant pneumococci. Acta Med Okayama, 2001 Jun, 55(3), 167 - 74 Genetic alteration of penicillin non-susceptible Streptococcus pneumoniae observed throughout recurrence of acute otitis media detected by amplified fragment length polymorphism analysis; Sugata K et al.; The prevalence of penicillin non-susceptible Streptococcus pneumoniae (PNSSP) is increasing among isolates from acute otitis media (AOM) . Repeated episodes of antibiotic exposure are a well-known risk factor for the isolation of PNSSP although otitis-prone or recurrent AOM cases frequently require repeated courses of antibiotic treatment . In order to evaluate the chronological alteration of S . pneumoniae during recurrences of AOM, strains of S . pneumoniae were isolated from 11 patients, each of whom had experienced 2-4 episodes of AOM, were examined . Every bacterial specimen obtained from a single episode of recurrent AOM was examined by PCR-based penicillin-binding protein (PBP) assay, serotyping, and amplified fragment length polymorphism (AFLP), then compared to other samples from the same case . Two cases (18.2%) showed strain diversity during repeated antibiotic treatments by serotyping or PBP-assay . By AFLP analysis, 6 cases (54.5%) demonstrated heterogeneous strains during recurrent AOM . Clonal survivors of previous episodes of AOM were not always the cause of subsequent episodes of AOM, even in otitis-prone cases. Nippon Ronen Igakkai Zasshi, 2001 May, 38(3), 281 - 4 {Infective endocarditis in the elderly}; Chikamori T et al.; To elucidate clinical features of infective endocarditis in the elderly, 20 elderly patients aged > or = 60 years were compared in detail with 30 others aged < 60 years retrospectively . Twelve of the 20 elderly patients had a calcific aortic valve or an artificial device as a predisposing heart disease, whereas 16 middle-aged patients had mitral valve prolapse or congenital heart disease (p = 0.001) . The prevalence of major extracardiac disorders such as neurological disease were higher in the elderly than in the middle (9/20 vs 3/30; p < 0.01) . The frequency of infected valve was similar; mitral in 8, aortic in 11 and other valves or congenital defect in 2 in the elderly versus 14, 11 and 6, respectively in the middle . Among 39 patients in whom causative microorganisms were identified, staphylococcus epidermidis was most frequently identified in the elderly (5/20), whereas streptococcus species was found in the middle (12/30) . Time from the onset of symptoms to correct diagnosis was usually delayed in the entire group; the delay was longer particularly in the elderly than in the middle-aged patients (72 +/- 87 vs 36 +/- 32 days; p < 0.1) . Maximal body temperature was less in the elderly than in the middle-aged patients (38.5 +/- 0.7 vs 39.3 +/- 1.1 degrees C; p < 0.01), whereas peak level of C-reactive protein (10.4 +/- 6.1 vs 13.0 +/- 7.9 mg/dL), the incidences of heart failure (9/20 vs 10/30), and embolic complications (7/20 vs 10/20) were similar in the 2 groups . Cardiac operation was performed less in the elderly than in the middle-aged patients (9/20 vs 21/30; p < 0.08) . Five elderly patients had disease-related mortality, whereas only one middle-aged patient died (p = 0.02) . These results suggest that although predisposing heart disease and causative microorganism in infective endocarditis are different between the elderly and middle-aged patients, the incidence of major complications are similar . However, due to the delay of correct diagnosis in the elderly who usually have major extracardiac disorders, the prognosis of infective endocarditis in the elderly is poor. Clin Oral Implants Res, 1998 Dec, 9(6), 365 - 73 C-telopeptide pyridinoline cross-links (ICTP) and periodontal pathogens associated with endosseous oral implants; Oringer RJ et al.; Detection of periodontal or peri-implant sites exhibiting progressing disease or those at risk of deterioration has proven difficult . Pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), a marker specific for bone degradation found in gingival crevicular fluid (GCF), has been associated with both bone and attachment loss in periodontitis and may be useful for predicting disease activity . The aim of this cross-sectional study was to examine the relationship between ICTP levels and subgingival species around implants and teeth from 20 partially and 2 fully edentulous patients . GCF and plaque samples were collected from the mesiobuccal site of each implant and tooth . Radioimmunoassay techniques were utilized to determine GCF ICTP levels . Plaque samples were analyzed utilizing checkerboard DNA-DNA hybridization . Traditional clinical parameters were assessed . Seventy-one implants and 370 teeth from 22 subjects were examined . ICTP levels and subgingival plaque composition were not significantly different between implants and teeth . Implant sites colonized by Prevotella intermedia, Capnocytophaga gingivalis, Fusobacterium nucleatum ss vincentii, and Streptococcus gordonii exhibited odds ratios of 12.4, 9.3, 8.1, and 6.7, respectively of detecting ICTP . These results suggest a relationship between elevated ICTP levels at implant sites and some species associated with disease progression . Longitudinal studies are necessary to determine whether elevated ICTP levels may predict the development of peri-implant bone loss. J Clin Microbiol, 2001 Jul, 39(7), 2719 - 21 Fluoroquinolone resistance is a poor surrogate marker for type II topoisomerase mutations in clinical isolates of Streptococcus pneumoniae; Millichap JJ et al.; The association between fluoroquinolone susceptibility and DNA mutations coding for amino acid substitutions in the quinolone resistance-determining region was assessed with 44 clinical isolates of Streptococcus pneumoniae . Twenty-three strains bore at least one amino acid substitution . Only seven strains with mutations were suggested by diminished susceptibility to ciprofloxacin (MIC, > or =2 microg/ml). J Clin Microbiol, 2001 Jul, 39(7), 2584 - 9 Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR; Riffon R et al.; Bovine mastitis is the most important source of loss for the dairy industry . A rapid and specific test for the detection of the main pathogens of bovine mastitis is not actually available . Molecular probes reacting in PCR with bacterial DNA from bovine milk, providing direct and rapid detection of Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus parauberis, and Streptococcus uberis, have been developed . Two sets of specific primers were designed for each of these microorganisms and appeared to discriminate close phylogenic bacterial species (e.g., S . agalactiae and S . dysgalactiae) . In addition, two sets of universal primers were designed to react as positive controls with all major pathogens of bovine mastitis . The sensitivities of the test using S . aureus DNA extracted from milk with and without a pre-PCR enzymatic lysis step of bacterial cells were compared . The detection limit of the assay was 3.125 x 10(2) CFU/ml of milk when S . aureus DNA was extracted with the pre-PCR enzymatic step compared to 5 x 10(3) CFU/ml of milk in the absence of the pre-PCR enzymatic step . This latter threshold of sensitivity is still compatible with its use as an efficient tool of diagnosis in bovine mastitis, allowing the elimination of expensive reagents . The two PCR tests avoid cumbersome and lengthy cultivation steps, can be performed within hours, and are sensitive, specific, and reliable for the direct detection in milk of the six most prevalent bacteria causing bovine mastitis. J Clin Microbiol, 2001 Jul, 39(7), 2565 - 71 Nomenclature of major antimicrobial-resistant clones of Streptococcus pneumoniae defined by the pneumococcal molecular epidemiology network; McGee L et al.; The emergence of disease caused by penicillin-resistant and multidrug-resistant pneumococci has become a global concern, necessitating the identification of the epidemiological spread of such strains . The Pneumococcal Molecular Epidemiology Network was established in 1997 under the auspices of the International Union of Microbiological Societies with the aim of characterizing, standardizing, naming, and classifying antimicrobial agent-resistant pneumococcal clones . Here we describe the nomenclature for 16 pneumococcal clones that have contributed to the increase in antimicrobial resistance worldwide . Guidelines for the recognition of these clones using molecular typing procedures (pulsed-field gel electrophoresis, BOX-PCR, and multilocus sequence typing) are presented, as are the penicillin-binding profiles and macrolide resistance determinants for the 16 clones . This network can serve as a prototype for the collaboration of scientists in identifying clones of important human pathogens and as a model for the development of other networks. Am J Epidemiol, 2001 Jul 1, 154(1), 85 - 92 Interpreting results from trials of pneumococcal conjugate vaccines: a statistical test for detecting vaccine-induced increases in carriage of nonvaccine serotypes; Lipsitch M; Conjugate vaccines against Streptococcus pneumoniae (pneumococcus) protect against nasopharyngeal carriage of serotypes included in the vaccine . However, in several clinical trials, vaccinees have shown increased carriage of nonvaccine serotypes of pneumococcus . These increases may be due to serotype replacement, if vaccine-induced protection against carriage of vaccine serotypes increases susceptibility to carriage of nonvaccine serotypes . Alternatively, observed increases may be an artifact of "unmasking," in which nonvaccine serotypes are more readily detected among vaccinees than among controls because vaccine serotypes are not present . In this paper, a statistical test for distinguishing serotype replacement from unmasking is described . The test attempts to reject a null model of unmasking alone; serotype replacement is inferred if the observed increase in detectable nonvaccine serotype carriage among vaccinees is significantly greater than that expected under the null model . Significance is assessed using the Bayesian "posterior predictive p value" as modified by Robins et al . (J Am Stat Assoc 2000;95:1143-56) . Analysis of data from a South African trial suggests that replacement may have occurred in the study, but results do not reach the conventional level of significance in rejecting the null hypothesis of unmasking (p = 0.074) . The author performs sensitivity analyses for the prior and for unmeasured confounding by differences in susceptibility to pneumococcus carriage . The implications of the findings and the assumptions and limitations of this technique are then discussed. FEMS Microbiol Lett, 2001 Jun 25, 200(2), 163 - 7 Transformation of an oral bacterium via chromosomal integration of free DNA in the presence of human saliva; Mercer DK et al.; Transformation of Streptococcus gordonii DL1 by free DNA was studied in human saliva . Competent S . gordonii could be transformed in vitro with plasmid DNA that had been taken into the human mouth . Transformation also occurred with a plasmid that cannot replicate in S . gordonii, but that has a region of chromosomal homology, by integration into the bacterial chromosome, although linearised plasmid DNA gave no transformants . Linear chromosomal DNA fragments did however transform S . gordonii/Tn916 efficiently in saliva when regions of homology with the recipient chromosome flanked the marker gene . These findings are discussed in relation to the potential for acquisition of DNA sequences, including genetically modified DNA, by gut and oral bacteria. FEMS Microbiol Lett, 2001 Jun 25, 200(2), 145 - 9 Inhibition of epithelial cell apoptosis by Porphyromonas gingivalis; Nakhjiri SF et al.; Porphyromonas gingivalis is periodontal pathogen that is capable of invading gingival epithelial cells (GECs) . Apoptotic responses of primary cultures of GECs to P . gingivalis were investigated with a DNA fragmentation ELISA assay . P . gingivalis induced a transient increase in GEC DNA fragmentation; however, after prolonged incubation GECs did not undergo apoptosis . Furthermore, P . gingivalis blocked apoptosis in GECs following stimulation with camptothecin . Immunoblotting of GECs with Bcl-2 or Bax antibodies showed that P . gingivalis up-regulated Bcl-2 levels in GECs, whereas Bax levels were transiently elevated and declined after 24 h stimulation . Streptococcus gordonii did not affect levels of either molecule . RT-PCR demonstrated that induction of Bcl-2 occurs at the transcriptional level . The results suggest that P . gingivalis can inhibit apoptosis in GECs by up-regulation of the anti-apoptotic molecule Bcl-2 . The prevention of host cell apoptosis may represent a strategy for P . gingivalis survival within invaded GECs. FEMS Microbiol Lett, 2001 Jun 25, 200(2), 137 - 43 Comparative genomics for identification of clone-specific sequence blocks in Streptococcus pneumoniae; Oggioni MR et al.; The partial genome sequences of a serotype 3 and a serotype 2 pneumococcal strain were compared to the complete type 4 pneumococcal genome . Over 500000 and 150000 base pairs of the partial genome data, obtained from published patents, were analysed respectively . Global alignment showed that nearly the whole genome is highly conserved in accordance with data of multilocus sequence typing of housekeeping genes . The search for clone-specific genes revealed 17 new open reading frames in the type 3 strain, while no new open reading frame was detected in the type 2 strain . Allelic variation of genes was restricted by the use of crude sequence data, but still permitted identification of some new alleles and the observation that all surface proteins present in the partial genome data were highly conserved . In both strains we observed also a variety of chromosomal rearrangements and variations due to mobile genetic elements . All together, this comparative genomic approach gives a genome-based overview of strain relatedness and a prospective on what could be expected when sequencing other pneumococcal strains. Acta Biomed Ateneo Parmense, 2000, 71 Suppl 1, 567 - 71 {Lavage of the birth canal with chlorhexidine: a new valid method for the prevention of perinatal infections}; Kaihura CT et al.; Perinatal morbidity and mortality are due to various infective agents, mainly represented by beta-hemolytic group B Streptococcus . The perinatal disease related to this infection is distinguished in Early-onset, characterised by pneumonia and sepsis, and Late-onset which leads to sepsis, meningitis and pneumonia . Various strategies were therefore proposed to prevent transmission including immunisation and chemoprophylaxis . All these methods however present adverse effects and are most of all expensive to carry out . Taha et al . reported an interesting experience regarding the reduction of perinatal infections following the cleansing of the birth canal with a solution of Chlorhexidine 0.25% during labour (1996-1997) . It seemed interesting for us to assess the applicability and efficacy of a new strategy of prophylaxis of perinatal infections in a Developing Country based on the association of two of the simple strategies proposed i.e.: cleansing the birth canal with chlorhexidine and chemoprophylaxis in cases with risk factors without culture screening . We studied two groups of patients: one in which cleansing of the birth canal was used and the second (control group) in which the old method already applied in the hospital (i.e . cleansing of the external genitals with Cetrimide 1%+ Chlorhexidine 0.1%) was carried out associated with antibiotic therapy when risk factors arose . We observed a total absence of neonatal mortality due to sepsis resulting from the association of the methods suggested even though the presence of sepsis evaluated through signs and symptoms like fever, poor feeding, apnoea or dyspnoea in newborns was similar in both groups. Can Vet J, 2001 Jun, 42(6), 465 - 7 Peritonitis in a llama caused by Streptococcus equi subsp . zooepidemicus; Hewson J et al.; A 7-month-old, male llama was diagnosed with peritonitis caused by Streptococcus equi subsp . zooepidemicus . Clinical findings, medical treatment, and case outcome are described . Hematogenous dissemination from suspected pneumonia is proposed as the route of infection in this case . Possible transmission of the organism through contact with horses is discussed. Kansenshogaku Zasshi, 2001 May, 75(5), 398 - 405 {A questionnaire survey on the theory of postoperative infection prophylaxis in orthopedics}; Shinagawa N et al.; A questionnaire survey on the theory of postoperative infection prophylaxis was conducted to obtain the consensus on perioperative antimicrobial use among orthopedists in Japan in the period from April to September 2000 . Fifty of the 91 orthopedists replied, and the following consensus was obtained . An antimicrobial prophylaxis (AMP) agent should be chosen based on their efficacy against the pathogens expected to be contaminants, such as Staphylococcus spp., and Streptococcus spp., Use an AMP agent that achieves a bactericidal concentration in both the serum and operating site . Use an AMP agent that has little unfavourable side effects . Use an AMP agent that affects minimally the normal bacterial flora . The most commonly used agents are the penicillins and first and second generation cephalosporins . The optimal strategy for most commonly used agents entails infusion of the first dose between approximately 30 minutes pre and post-skin incision and the therapeutic levels should be maintained throughout the operation . The AMP agents having no cross-resistance to the prophylactic agents should be used, if postoperative infection is suspected or developed . The most commonly used agent for both clean operations with or without foreign implants and dirty operations is cefazolin (CEZ), followed by cefotiam (CTM) and flomoxef (FMOX). J Infect Dis, 2001 Jul 15, 184(2), 166 - 73 Epub 2001 Jun 15. Enhanced susceptibility to superantigen-associated streptococcal sepsis in human leukocyte antigen-DQ transgenic mice; Sriskandan S et al.; Bacterial superantigens are believed to cause septic shock, although, because of the lack of superantigen-sensitive infection models, proof that superantigenicity underlies shock pathogenesis is lacking . This work demonstrates a clear superantigen effect in septic shock resulting from bacterial infection . Transgenic expression of human leukocyte antigen (HLA)-DQ, but not HLA-DR, specifically augments lymphocyte responses to streptococcal pyrogenic exotoxin A (SPEA) . HLA-DQ transgenic mice had increased mortality after administration of SPEA or infection with Streptococcus pyogenes . Immune activation during infection was HLA-DQ transgene-dependent and was manifested by Vbeta-specific T cell repertoire changes and widespread lymphoblastic tissue infiltration . Unlike earlier models, which used toxin-induced shock, these T cell superantigen responses and lymphoblastoid changes were observed during invasive streptococcal sepsis . Lymphoid activation was undetectable in HLA-DQ mice infected with an isogenic SPEA(-) strain, which proves that a single superantigen can play a role in sepsis pathogenesis. Vet Microbiol, 2001 Sep 3, 82(1), 69 - 80 Experimental airborne transmission of Streptococcus suis capsular type 2 in pigs; Berthelot-Herault F et al.; Experimental airborne transmission of Streptococcus suis type 2 was studied in specific pathogen free piglets . Forty piglets were allotted to five groups of eight 7-week-old animals and housed in three separated units . Negative control pigs (group 1) were housed in unit A and infected batches were housed in units B (group 2) and C (groups 4) . In units B and C, non-inoculated groups (groups 3 and 5, respectively), 40 cm distant from the respective inoculated group and without any physical contact between them, also took place . Six animals of groups 2 and 4 were inoculated intravenously with 2 x 10(8) colony forming units (cfu) of a mild and a high virulent S . suis strains, respectively . The remaining animals in these groups and pigs from groups 1, 3, 5 received broth medium in the same way . Differences among virulence of S . suis capsular type 2 were observed in inoculated pigs of groups 2 and 4 . Pigs from group 2 became carriers, showing only mild symptoms . By contrast, animals from group 4 presented an acute form of the disease . All the indirect contact pigs in groups 3 and 5 had S . suis in palatine tonsils from day 6 after the infection and they presented clinical manifestations similar to those observed in experimentally infected pigs . Two direct contact animals were also contaminated in the upper respiratory tract but surprisingly they did not show any symptoms . Airborne transmission of S . suis in experimentally pigs was demonstrated in the present study . Indirect infections, as described in this study, are a more realistic way to infect pigs than other experimental procedures and may be used to further study the pathogenesis of the infection caused by this important pathogen. Carbohydr Res, 2001 Jun 22, 333(1), 19 - 26 Dextran acceptor reaction of Streptococcus sobrinus glucosyltransferase GTF-I as revealed by using uniformly 13C-labeled sucrose; Mukasa H et al.; A sucrose glucosyltransferase GTF-I from cariogenic Streptococcus sobrinus transferred the uniformly 13C-labeled glucosyl residue ({U-(13)C}Glc) from {U-(13)C}sucrose to exogenous dextran T500 at the non-reducing-end, mostly by alpha-(1-->6) linkages and partially by alpha-(1-->3) linkages, as revealed by the 13C-(13)C NMR coupling pattern . With increasing amounts of {U-(13)C}sucrose, transfer of {U-(13)C}Glc to the alpha-(1-->3)-linked chain became predominant without increase in the number of chains . The transfer of {U-(13)C}Glc to an isomaltopentaose acceptor occurred similarly to its transfer to T500 . alpha-(1-->3)-branches in the {U-(13)C}dextran, specifically synthesized from {U-(13)C}sucrose by a Streptococcus bovis dextransucrase, were not formed by GTF-I, as judged by the observation that a newly-formed alpha-1,3,6-branched {U-(13)C}Glc was not detected, which could have been formed by transferring the unlabeled Glc from sucrose to the internal alpha-(1-->6)-linked {U-(13)C}Glc at C-3 . The 13C-(13)C one-bond coupling constants (1J) were also recorded for the C-1--C-6 bond of the internal alpha-(1-->6)-linked {U-(13)C}Glc and of the non-reducing-end {U-(13)C}Glc. Glycoconj J, 2000 Jul-Sep, 17(7-9), 659 - 64 Safe as mother's milk: carbohydrates as future anti-adhesion drugs for bacterial diseases; Sharon N et al.; The majority of infectious diseases are initiated by adhesion of pathogenic organisms to the tissues of the host . In many cases, this adhesion is mediated by lectins present on the surface of the infectious organism that bind to complementary carbohydrates on the surface of the host tissues . Lectin-deficient mutants often lack ability to initiate infection . Soluble carbohydrates recognized by the bacterial lectins block the adhesion of the bacteria to animal cells in vitro . Moreover, they have also been shown to protect against experimental infection by lectin-carrying bacteria in different organs of mammals such as mice, rabbits, calves and monkeys . In a phase II clinical trial, a pentasaccharide shown to have anti-adhesive activity against Streptococcus pneumoniae and Hemophilus influenzae in vitro failed to protect young children from nasopharyngeal colonization with these organisms and from developing otitis media . This could be because insufficient drug was delivered via nasal spray, because bacteria express multiple specificities, the inhibition of which may require a cocktail of oligosaccharides, or because children have different carbohydrate receptors from those of adults . The results of a clinical trial in which N-acetylneuraminyl(alpha2-3)lactose was administered orally to Helicobacter pylori positive patients in an effort to reduce or eradicate bacterial colonization, are awaited with interest . Although the high cost of production of the required oligosaccharides is falling with the recent introduction of enzymatic methods of synthesis, new technologies, in particular the use of engineered bacteria, promise to lower it even further . Attachment of the oligosaccharides to soluble polymeric carriers will increase greatly their effectiveness as antiadhesion agents . There is no doubt that anti-adhesive oligosaccharides will in the near future join the arsenal of drugs for the therapy of bacterial diseases. Respir Med, 2001 Jun, 95 Suppl A, S5 - 11; discussion S26-7 The hidden impact of antibacterial resistance in respiratory tract infection . Clinical failures: the tip of the iceberg? Garau J. In pneumococcal meningitis, it is well accepted that resistance in Streptococcus pneumoniae compromises clinical outcome . However, the clinical impact of increasing resistance on community-acquired respiratory tract infections (RTIs) is less clear . Bacteriological eradication should be the aim of antimicrobial therapy . The pharmacodynamics (potency and pharmacokinetics) of an antimicrobial agent against the infecting pathogen can be used to predict the potential for bacterial eradication . Surveillance of clinical isolates from community-acquired RTIs shows that, in many countries, there is a trend towards an increasing prevalence of drug-resistant S . pneumoniae . Results from a number of published clinical trials suggest that resistance has not compromised the clinical efficacy of aminopenicillins when used at the correct dose . However, emerging data indicate that resistance is compromising the efficacy of some other routinely used antimicrobials . There are reports of clinical and bacteriological failure with macrolides and fluoroquinolones in patients with community-acquired pneumonia . Recent retrospective analyses and increasing sporadic reports of clinical failure with these agents may be more representative of the true situation . These reports suggest a need to reassess current empirical therapeutic recommendations for the treatment of community-acquired RTIs. Respir Med, 2001 Jun, 95 Suppl A, S12 - 9; discussion S26-7 The hidden impact of antibacterial resistance in respiratory tract infection . Re-evaluating current antibiotic therapy; Craig WA; Pharmacokinetic/pharmacodynamic (PK/PD) parameters derived from animal and clinical models of infection are used to predict bacteriological efficacy . Growing evidence from the clinical setting supports the validity of these parameters in guiding antimicrobial therapy . For example, in otitis media and sinusitis, high bacteriological cure rates are obtained when serum concentrations of beta-lactams and macrolides exceed the MIC of the infecting pathogen for at least 40% of the dosing interval . Likewise, the 24-hour AUC/MIC ratio is a good predictor of both bacteriological and clinical efficacy for azithromycin in otitis media and fluoroquinolones in bacterial pneumonia . The value of PK/PD relationships has been recognized by the National Committee for Clinical Laboratory Standards (NCCLS) as another important factor to consider when establishing susceptibility breakpoints . Recent changes to NCCLS breakpoints for oral beta-lactams for Streptococcus pneumoniae reflect this . Also, PK/PD parameters may play a role in predicting the impact of an antibiotic on the development and spread of resistant organisms . In an era of increasing resistance, we should select agents and doses that provide drug concentrations that exceed the magnitude of the PK/PD parameter required both for efficacy and to combat the emergence and spread of bacterial resistance. Biosens Bioelectron, 2000, 15(9-10), 511 - 4 Immobilization of pneumococcal polysaccharide vaccine on silicon oxide wafer for an acoustical biosensor; Dutra RF et al.; One the most important aspects of a biosensor is related to immobilization and maintenance of specific reference compounds on sensing surfaces . A method for the immobilization of polysaccharides to a silicon oxide surface intended for Surface Acoustical Waves (SAW) sensors is described . Silicon oxide is a hydrophobic inorganic support used for the fabrication of many electronic devices . The pneumococcal polysaccharide (PPS) vaccine is immobilized via Protein A after pre-treatment of the surface with hydrochloric acid . The effects of non-specific binding are discussed . The results indicate that the immobilization of PPS via Protein A increases the sensitivity of detecting Streptococcus pneumoniae antibodies in human sera and offers greater reproducibility of response compared with ELISA methods . The principles of this technique are simple and are applicable to the immobilization of many capsular polysaccharides. Pediatr Infect Dis J, 2001 Jun, 20(6), 607 - 11 Bacterial etiology of acute myringitis in children less than two years of age; Palmu AA et al.; BACKGROUND: Acute myringitis is an inflammation of the tympanic membrane that occurs alone or in association with external otitis or otitis media . The two clinical entities, based on physical examination, are bullous myringitis and hemorrhagic myringitis . OBJECTIVES: To investigate the association of concomitant middle ear disease with acute myringitis and to analyze the bacteriologic findings of the middle ear fluid in children with acute myringitis . METHODS: A prospective longitudinal cohort study of 2028 children age 7 to 24 months at primary care level in the Finnish Otitis Media Vaccine Trial . Matched case-control design for analysis of bacterial pathogen distribution . RESULTS: There were 82 children in whom 92 ears were diagnosed with acute bullous myringitis and 37 children in whom 40 ears were diagnosed with hemorrhagic myringitis during the follow-up . Middle ear disease was associated with bullous myringitis in 97% of ears and with hemorrhagic myringitis in 82% of ears . Bacterial pathogen distribution was similar to that of acute otitis media, although a higher proportion of Streptococcus pneumoniae was detected in both bullous and hemorrhagic acute myringitis . CONCLUSIONS: Middle ear fluid was present in vast majority of ears with acute myringitis in young children . The same etiologic bacteria were found in acute myringitis as in acute otitis media, but S . pneumoniae was the major pathogen . Acute bullous myringitis should be treated as acute otitis media in children <2 years of age. J Antimicrob Chemother, 2001 Jul, 48(1), 29 - 36 Activity of non-fluorinated quinolones (NFQs) against quinolone-resistant Escherichia coli and Streptococcus pneumoniae; Roychoudhury S et al.; The newly developed 8-methoxy, non-fluorinated quinolones (NFQs) were studied to elucidate their enzyme inhibitory activity against wild-type and mutant GyrA (Ser-83-->Trp) forms of Escherichia coli DNA gyrase . Using a DNA supercoiling inhibition assay, the NFQs were found to inhibit 50% (IC50) of the E . coli DNA gyrase activity in the 1.6-3.2 mg/L concentration range and were comparable to ciprofloxacin . However, against the GyrA (Ser-83-->Trp) mutant, the NFQs were approximately 16-fold more potent than ciprofloxacin . Antibacterial potency of the NFQs was investigated using clinical isolates of E . coli and penicillin-resistant Streptococcus pneumoniae (PRSP), including strains with reduced susceptibility to quinolones . Against 20 uncharacterized clinical isolates of E . coli, the MIC90s of the NFQs were in the 0.125-0.25 mg/L range while those of ciprofloxacin, trovafloxacin, gatifloxacin and clinafloxacin were in the 0.016-0.125 mg/L range . Against clinical isolates with characterized mutations in gyrA and parC, PGE9262932, an NFQ, was two- to eight-fold more potent than ciprofloxacin . Against 23 clinical isolates of PRSP, the NFQs (MIC90 0.031-0.125 mg/L) were more potent than ciprofloxacin, trovafloxacin, and gatifloxacin (MIC90 0.25-2.0 mg/L), and at least as potent as clinafloxacin (MIC90 0.125 mg/L) . Against S . pneumoniae strains with gyrA and parC mutations, the NFQs (MIC 0.125-1.0 mg/L) were more potent than ciprofloxacin, trovafloxacin and gatifloxacin (MIC 4-32 mg/L), and comparable to clinafloxacin (MIC 0.5-1 mg/L). J Antimicrob Chemother, 2001 Jul, 48(1), 15 - 22 The effect of pharmacokinetics on the bactericidal activity of ciprofloxacin and sparfloxacin against Streptococcus pneumoniae and the emergence of resistance; Thorburn CE et al.; The pharmacokinetics of ciprofloxacin and sparfloxacin were simulated in vitro and the effects of pharmacodynamic parameters on bactericidal activity and the emergence of quinolone resistance were examined for Streptococcus pneumoniae . Simulated serum concentrations of ciprofloxacin 500 mg bd were more rapidly bactericidal than sparfloxacin 200 mg bd, despite lower values for the key pharmacodynamic parameters (AUC/MIC and C(max)/MIC) . This was possibly related to the slower oral absorption of sparfloxacin, which delayed achievement of the MIC compared with ciprofloxacin . In addition, sparfloxacin was shown to have similar bactericidal activity to ciprofloxacin when tested at the same concentrations, despite its four-fold better potency in MIC terms . The emergence of resistance following exposure to ciprofloxacin appeared to be dependent on the C(max)/MIC ratio and the AUC above the MIC, but not the AUC/MIC ratio . Resistance (at least four-fold increase in MIC) developed when the C(max)/MIC ratio was less than four or the AUC above the MIC was less than 10, and the resulting cultures regrew fully . In contrast, pneumococci with a two- to four-fold increase in sparfloxacin MIC were selected in the presence of serum concentrations of sparfloxacin despite a C(max)/MIC ratio higher than 12, but these isolates remained clinically susceptible by breakpoint MIC and their growth was inhibited by repeated dosage of sparfloxacin . Nevertheless, the selection of pneumococci with reduced susceptibility, and the possibility of further mutation to highly resistant strains supports the use of quinolones that rapidly eradicate pneumococci at conventional doses and achieve concentrations, in both serum and tissues, which exceed at least 4 x MIC. Immunopharmacol Immunotoxicol, 2001 May, 23(2), 153 - 61 Splenectomy and sepsis: the role of the spleen in the immune-mediated bacterial clearance; Altamura M et al.; Over the past few years, many observations of overwhelming post splenectomy bacterial infections have been reported . Streptococcus pneumoniae is the aetiologic agent in about 80% of cases, but also gram-negative bacteria are involved in the development of fatal infections in splenectomized patients . Functionally, the spleen plays a fundamental role in bacterial clearance either by antibody response or macrophage bactericidal capacity . At the same time, there is evidence that the spleen also contributes to bacterial endotoxin detoxification . Finally, the mechanisms responsible for gram-positive and gram-negative sepsis in the splenectomized host and possible therapeutical approaches able to neutralize bacterial products endowed with noxious effects are discussed. J Dairy Sci, 2001 Jun, 84(6), 1545 - 9 Efficacy of a new premilking teat disinfectant containing a phenolic combination for the prevention of mastitis; Oliver SP et al.; A teat disinfectant containing a phenolic combination was evaluated in a natural exposure study in two dairy research herds . Premilking teat disinfection was compared with a negative control using a split-udder experimental design . In both herds, premilking and postmilking teat disinfections with the phenolic combination were significantly more effective in preventing new intramammary infection (IMI) than was postmilking teat disinfection only . Clinical mastitis and new IMI by Streptococcus uberis, Streptococcus dysgalactiae, Gram-negative pathogens, and coagulase-negative Staphylococcus species were significantly lower in quarters of cows with teats predipped and postdipped than in quarters with teats postdipped only . No chapping or teat skin irritation was observed . Premilking teat disinfection with the phenolic combination in association with good udder preparation and postmilking teat disinfection can further reduce the occurrence of new IMI by numerous mastitis pathogens during lactation. Klin Padiatr, 2001 May-Jun, 213(3), 109 - 13 {Value of pneumococcal vaccination in infants and young children}; Adam D et al.; Streptococcus pneumoniae is a common cause of meningitis, sepsis, pneumonia, acute otitis media, and sinusitis in children . Children younger than 24 months have the highest rates of invasive pneumococcal infections (Germany 1997-1999: 19.5/100,000/year) . Pneumococcal infections cause in Germany 220-250 cases of meningitis, about 50,000 of pneumonia (children younger than 5 years) and more than 1 million cases of otitis media (children) annually . The case-fatality rate for invasive pneumococcal diseases is high (1997-1999 5.5%, meningitis 8.3%) . 20-30% of survivors from meningitis suffer from CNS-related sequelea . In children up to 2 years vaccination with the heptavalent pneumococcal conjugate vaccine can reduce invasive pneumococcal diseases by about 80% and otitis media and recurrent otitis media by 6% and 9-16%, respectively . Due to the increased risk of pneumococcal infections in the first two years of live all children of this age group should be vaccinated . The high rate of resistance of pneumococci against macrolides in Germany, the high rate of non-licensed antibiotics in infants and the inefficacy of the 23-valent vaccine in children younger than 2 years makes the new vaccine to a necessary alternative. Proc Natl Acad Sci U S A, 2001 Jun 19, 98(13), 7552 - 7 Multilocus analysis of extracellular putative virulence proteins made by group A Streptococcus: population genetics, human serologic response, and gene transcription; Reid SD et al.; Species of pathogenic microbes are composed of an array of evolutionarily distinct chromosomal genotypes characterized by diversity in gene content and sequence (allelic variation) . The occurrence of substantial genetic diversity has hindered progress in developing a comprehensive understanding of the molecular basis of virulence and new therapeutics such as vaccines . To provide new information that bears on these issues, 11 genes encoding extracellular proteins in the human bacterial pathogen group A Streptococcus identified by analysis of four genomes were studied . Eight of the 11 genes encode proteins with a LPXTG(L) motif that covalently links Gram-positive virulence factors to the bacterial cell surface . Sequence analysis of the 11 genes in 37 geographically and phylogenetically diverse group A Streptococcus strains cultured from patients with different infection types found that recent horizontal gene transfer has contributed substantially to chromosomal diversity . Regions of the inferred proteins likely to interact with the host were identified by molecular population genetic analysis, and Western immunoblot analysis with sera from infected patients confirmed that they were antigenic . Real-time reverse transcriptase-PCR (TaqMan) assays found that transcription of six of the 11 genes was substantially up-regulated in the stationary phase . In addition, transcription of many genes was influenced by the covR and mga trans-acting gene regulatory loci . Multilocus investigation of putative virulence genes by the integrated approach described herein provides an important strategy to aid microbial pathogenesis research and rapidly identify new targets for therapeutics research. Brain Pathol, 2001 Jul, 11(3), 282 - 95 Transcriptional regulation of caspases in experimental pneumococcal meningitis; von Mering M et al.; Apoptosis and necrosis in brain account for neurological sequelae in survivors of bacterial meningitis . In meningitis, several mechanisms may trigger death pathways leading to activation of transcription factors regulating caspases mRNA synthesis . Therefore, we used a multiprobe RNA protection assay (RPA) to examine the expression of 9 caspase-mRNA in the course of experimental Streptococcus pneumoniae meningitis in mouse brain . Caspase-6, -7 and -11 mRNA were elevated 6 hours after infection . 12 hours after infection caspases-1, -2, -8 and -12 mRNA rose . Caspase-14 mRNA was elevated 18 h and caspase-3 mRNA 24 h after infection . In situ hybridization detected caspases-3, -8, -11 and -12 mRNA in neurons of the hippocampal formation and neocortex . Development of sepsis was paralleled by increased transcription of caspases mRNA in the spleen . In TNFalpha-deficient mice all caspases examined were less upregulated, in TNF-receptor 1/2 knockout mice caspases-1, -2, -7, -11 and -14 mRNA were increased compared to infected control animals . In caspase-1 deficient mice, caspases-11, and -12 mRNA levels did not rise in meningitis indicating the necessity of caspase-1 activating these caspases . Hippocampal formations of newborn mice incubated with heat-inactivated S . pneumoniae R6 showed upregulation of caspase-1, -3, -11 and -12 mRNA . These observations suggest a tightly regulated caspases network at the transcriptional level in addition to the known cascade at the protein level. Southeast Asian J Trop Med Public Health, 2000 Dec, 31(4), 679 - 83 Risk factors for penicillin-resistant Streptococcus pneumoniae acquisition in patients in Bangkok; Dejthevapor C et al.; To identify risk factors for acquisition of penicillin-resistant Streptococcus pneumoniae (PRSP) in patients in Bangkok, using a case-control study, the study included patients with clinical specimens which grew S . pneumoniae during January to December 1997, treated at a teaching hospital in Bangkok . Penicillin susceptibility was determined by E-test and strains with MIC of > 0.1 microg/ml were considered resistant . Cases were the patients who had PRSP, and patients who had penicillin-susceptible S . pneumoniae (PSSP) were controls . The study variables included age 15 years or younger, immunocompromised status, ventilatory support, and antibiotic use or hospitalization within the previous 3 months . There were 73 cases and 51 controls . Their ages were 0 to 87 years, with median age of cases 4 and controls 49 years . Pneumonia was the most common type of infection, being 47% in cases and 45% in controls . Univariate analysis revealed significant association of PRSP acquisition with previous antibiotic use (p<0.0001), age < or = 15 years (p=0.001) and previous hospitalization (p=0.002) . Logistic regression analysis in order to adjust for confounding effects showed that the only significant risk factor was previous antibiotic use (OR 18.4; 95% CI 6.2-54.6) . The major risk factor for acquisition of PRSP in this study population is recent antibiotic use . Decreased antibiotic use would reduce risk of acquisition of PRSP. Rev Med Chil, 2001 Apr, 129(4), 397 - 404 {Complications in children with varicella in 4 hospitals in Santiago, Chile: clinical spectrum and estimation of direct costs}; Abarca K et al.; BACKGROUND: The knowledge of varicella complications and their associated cost may help for a better evaluation of varicella immunization benefits . AIM: To determine frequency, type, outcome and affected population of varicella complications in children requiring hospitalization, and to estimate their direct costs . MATERIAL AND METHODS: Retrospective analysis of medical records of children admitted to four hospitals in Santiago, Chile, due to varicella complications between January 1997 and February 1999 . Calculation of direct costs of hospitalizations in a sample of 30 patients . RESULTS: One hundred fifty four patients were identified, 74% were younger than 5 years old, only one was immunocompromised . Complications identified were skin and soft tissue infections in 63%, invasive infections in 25.3%, neurological in 7.1% and miscellaneous in 4.5% . Staphylococcus aureus and Group A beta-haemolytic Streptococcus (GABS) were predominantly isolated . S . aureus was the main agent identified in superficial infections and GABS in invasive infections (sterile sites) . Two patients died due to invasive infections (streptococcal toxic shock and S . aureus septicaemia) and 11 required surgical procedures . The average cost per hospitalization was US$ 600 in public hospitals and US$ 1,800 in the private hospital . CONCLUSIONS: Varicella complications requiring hospitalization are due mainly to bacterial infections and they affect immunocompetent toddlers . These complications can be severe and even fatal. J Laryngol Otol, 2001 May, 115(5), 363 - 8 Nebulized surfactant as a treatment choice for otitis media with effusion: an experimental study in the rabbit; Koten M et al.; Exogenous surfactant can improve eustachian tube function in experimentally induced otitis media with effusion (OME) . Performing tympanometric recordings, the efficacy of inhaled nebulized surfactant, as compared with inhaled nebulized physiological saline was investigated, for the treatment of OME experimentally induced in the rabbit by intrabullar inoculation of heat-killed Streptococcus pneumoniae . In addition, the histological changes in middle ears after the treatment were investigated in order to establish whether the pathological findings correlated with the results . Middle-ear pressure values before, and after, treatment were analyzed by the Wilcoxon statistical method, and the Mann-Whitney U test was used to compare the post-treatment values between groups . In all ears with OME in the affected animals, which were treated with nebulized surfactant inhalation, a positively significant (p<0.05) increase of pressure more than 20 daPa was recorded . In the control group, after inhalation of nebulized physiological saline, there was no positive increase in the affected middle-ear pressures; on the contrary, more negative pressure changes were recorded . In the histological evaluation, middle-ear epithelia and sub-epithelial space were normal in surfactant-treated ears with OME, whereas mucosal thickening with an oedematous sub-epithelial space containing occasional inflammatory cells and increases in connective tissue and vascularity, and effusions on the epithelial surface were present in the ears with OME in the control group . The significant improvement in the negative middle-ear pressure after nebulized surfactant treatment and the histological findings shown in our study can support the theory that surface-active agents are of importance in eustachian tube function even under pathologic conditions, such as OME. Ann Pharmacother, 2001 Jun, 35(6), 687 - 90 Levofloxacin failure in a patient with pneumococcal pneumonia; Empey PE et al.; OBJECTIVE: To report a case of levofloxacin failure in a patient with a penicillin-sensitive Streptococcus pneumoniae pneumonia . CASE SUMMARY: A previously healthy, immunocompetent, 53-year-old white man presented with penicillin-sensitive S . pneumoniae pneumonia . The patient was empirically placed on levofloxacin monotherapy, which was continued due to a local penicillin shortage . When the patient failed to improve, further susceptibility testing was ordered . The organism was found to have a penicillin minimum inhibitory concentration (MIC) of 0.023 microgram/mL and a levofloxacin MIC of 6 micrograms/mL . Effective antimicrobial therapy was delayed, as clinicians did not anticipate fluoroquinolone resistance . DISCUSSION: Newer fluoroquinolones such as levofloxacin have good activity against most S . pneumoniae isolates and are used for the treatment of pneumonia . Although resistance to these agents is rare, it has been reported . Current guidelines from the National Committee for Clinical Laboratory Standards do not recommend initial fluoroquinolone susceptibility testing . CONCLUSIONS: As fluoroquinolone resistance may not be identified by susceptibility patterns to other antibiotics, early fluoroquinolone susceptibility testing and increased awareness of resistance may aid clinicians in their treatment of pneumococcal disease. J Biol Chem, 2001 Aug 31, 276(35), 33121 - 8 Epub 2001 Jun 18. Characterization of selected strains of pneumococcal surface protein A; Jedrzejas MJ et al.; Several proteins, in addition to the polysaccharide capsule, have recently been implicated in the full virulence of the Streptococcus pneumoniae bacterial pathogen . One of these novel virulence factors of S . pneumoniae is pneumococcal surface protein A (PspA) . The N-terminal, cell surface exposed, and functional part of PspA is essential for full pneumococcal virulence, as evidenced by the fact that antibodies raised against this part of the protein are protective against pneumococcal infections . PspA has recently been implicated in anti-complementary function as it reduces complement-mediated clearance and phagocytosis of pneumococci . Several recombinant N-terminal fragments of PspA from different strains of pneumococci, Rx1, BG9739, BG6380, EF3296, and EF5668, were analyzed using circular dichroism, analytical ultracentrifugation sedimentation velocity and equilibrium methods, and sequence homology . Uniformly, all strains of PspA molecules studied have a high alpha-helical secondary structure content and they adopt predominantly a coiled-coil structure with an elongated, likely rod-like shape . No beta-sheet structures were detected for any of the PspA molecules analyzed . All PspAs were found to be monomeric in solution with the exception of the BG9739 strain which had the propensity to partially aggregate but only into a tetrameric form . These structural properties were correlated with the functional, anti-complementary properties of PspA molecules based on the polar distribution of highly charged termini of its coiled-coil domain . The recombinant Rx1 PspA is currently under consideration for pneumococcal vaccine development. J Microbiol Methods, 2001 Aug, 46(2), 107 - 17 Isolation of human plasma-inducible, growth phase- and temperature-regulated gene fusions in Streptococcus pyogenes using a Tn917-lacZ transposon; Sledjeski DD et al.; Streptococcus pyogenes is capable of causing a variety of human diseases ranging from superficial or deep tissue infections to non-infectious post-streptococcal infection sequelae . In this paper, we report the use of a Tn917-lacZ transposon to isolate random lacZ transcription fusions in the S . pyogenes chromosome . Libraries of random Tn917-lacZ mutants were generated in a representative opacity factor positive strain CS101 (M49) and an opacity factor negative strain 1881 (M1) . Several different mutant phenotypes were isolated . These included: temperature-regulated promoters, growth phase/cell density-regulated promoters and a human plasma-inducible promoter . Expression of the temperature-regulated fusions was 5-10-fold higher when grown at 30 degrees C compared to growth at 37 degrees C . The growth phase-regulated fusions were induced 30-fold at late exponential phase and were repressed by a diffusible S . pyogenes factor(s) . Expression of the human plasma-inducible fusion was induced 10-15-fold by human plasma or sera, 4-fold by rabbit sera and was repressed by horse and mouse sera . In addition, hemolysin negative and capsule over expression mutants were isolated . These results demonstrate the utility of Tn917-lacZ mutagenesis for the identification of S . pyogenes promoters. J Microbiol Methods, 2001 Aug, 46(2), 87 - 97 Use of surface-enhanced laser desorption ionization protein chip system to analyze streptococcal exotoxin B activity secreted by Streptococcus pyogenes; Boyle MD et al.; Ciphergen surface-enhanced laser desorption ionization (SELDI) protein chip technology was used to analyze the secretion and autoactivation of the Streptococcus pyogenes cysteine protease SpeB . This method allowed rapid identification of both the zymogen form of the protein Mr approximately 41,000 and the fully active enzyme Mr approximately 28,500 . SpeB production in culture supernatants was demonstrated to be growth-phase regulated and SpeB positive and negative variants of a blood passaged S . pyogenes isolate could readily be distinguished . In kinetic studies of the autoactivation of the zymogen form of SpeB, the sequential generation of four intermediates was detected before the accumulation of the fully active enzyme . The methods described enabled enhanced speed, use of lower sample volumes and concentrations, and a more complete molecular characterization of SpeB than allowed by existing methods of analysis using SDS-PAGE and Western immunoblotting. Int J Syst Evol Microbiol, 2001 May, 51(Pt 3), 1147 - 50 Streptococcus ovis sp . nov., isolated from sheep; Collins MD et al.; Seven strains of an unknown Gram-positive catalase-negative chain-forming coccus-shaped organism isolated from clinical specimens from sheep were characterized by phenotypic and molecular taxonomic methods . Comparative 16S rRNA gene sequencing studies demonstrated that the bacterium represents a new sub-line within the genus Streptococcus . The unknown bacterium was readily distinguished from recognized streptococcal species by biochemical tests and electrophoretic analysis of whole-cell proteins . Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Streptococcus ovis sp . nov . The type strain of Streptococcus ovis is CCUG 39485T (= LMG 19174T). EMBO J, 2001 Jun 15, 20(12), 3046 - 55 EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG; Collin M et al.; Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors . In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma . This activity is associated with the secretion of a novel 108 kDa protein denoted EndoS . EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface . EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG . In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner . This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. Arch Otolaryngol Head Neck Surg, 2001 Jun, 127(6), 650 - 4 Streptococcus milleri: an organism for head and neck infections and abscess; Han JK et al.; BACKGROUND: Streptococcus milleri, a commensal organism, has the potential to cause significant morbidity . There is a paucity of published data regarding this organism in the head and neck . OBJECTIVES: To identify and assess the presentation, treatment, and outcomes of pediatric patients affected by this pathogen . STUDY DESIGN: Review of the Department of Pathology database at Children's Hospital of Wisconsin, Milwaukee, between 1997 and 1999 identified 26 patients with cultures positive for S milleri group (SMG) bacteria . Retrospective chart analysis examined the demographic data, site of origin of infection, additional organisms cultured, symptoms, treatments, and complications . RESULTS: Sixteen patients had SMG infections involving the head and neck region . Sites of origin included the paranasal sinuses, dental, facial soft tissues, deep neck spaces, peritonsillar region, and a tracheostomy site . The paranasal sinuses were the most common site in 37% (6/16) . Streptococcus milleri was the only isolate in 69% (11) of the infections . Significant local extension occurred in 56% (9/16) of the patients and included the orbit, skull base, cranium, and deep neck spaces . All patients had surgical drainage and 15 also received intravenous antibiotic treatment . One complication of osteomyelitis of the frontal bone occurred with resolution after surgical debridement and intravenous antibiotic treatment . CONCLUSIONS: Streptococcus milleri can be an aggressive pathogen in the head and neck with a propensity for abscess formation and local extension of the infection in a pediatric population . Surgical drainage with antibiotics is generally successful in management of the condition . However, emerging penicillin resistance and the ability for local extension require suspicion of incomplete treatment if clinical symptoms persist.
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