Streptococcal

Vaccine, 2000 Dec 8, 19(9-10), 1159 - 66
Serotype of Streptococcus pneumoniae capsular polysaccharide can modify the Th1/Th2 cytokine profile and IgG subclass response to pneumococal-CRM(197) conjugate vaccines in a murine model; Mawas F et al.; The cellular and antibody responses to type 14 and type 19F Streptococcus pneumoniae capsular polysaccharides (PS) conjugated to CRM(197) were investigated in a mouse model developed for pre-clinical evaluation and quality control of pneumococcal conjugate vaccines . Total IgG antibody and IgG subclasses against PS and the carrier protein for both conjugates were measured in addition to the T cell proliferation and cytokine profiles induced by these conjugates . While unconjugated PS 14 and 19F were at best only weakly immunogenic, both types of conjugate induced strong primary and secondary IgG responses to PS . The responses induced by the two conjugates to the carrier protein were very different; a high level of anti-CRM(197) IgG was induced only by the PS19F conjugate whereas a very weak response was induced by the PS14 conjugate . Interestingly, the IgG subclass distribution was different for the two conjugates; for PS19F conjugate, the IgG response was almost completely of IgG1 subclass with low levels of IgG3 and IgG2a while the response to PS14 conjugate was mainly of the IgG1 and IgG2a subclasses with a low level of IgG3 . The anti-CRM(197) IgG subclass distribution was identical with that to the corresponding conjugated PS . Both types of conjugate induced strong T cell proliferation to recall antigens but induced different patterns of cytokine response in immune spleen cells which were indicative of a Th0 response or a mixture of Th1 and Th2 responses with a bias towards Th2 response in PS19F-CRM(197) immunised mice . In conclusion, PS14- and PS19F-CRM(197) conjugates induced different IgG subclass patterns as a result of inducing different patterns of cytokine response to the carrier protein . This indicates that the serotype of PS can modify the Th1/Th2 response to the carrier protein, which has a direct effect and can predict the IgG subclass of the PS response . Finally, we conclude that this model appears suitable for studying the immunogenicity and immune interaction of different components of multivalent pneumococcal conjugate vaccines and may be applicable to their pre-clinical evaluation and quality control.

Microbes Infect, 2000 Nov, 2(14), 1733 - 42
Pathogenesis of neonatal Streptococcus agalactiae infections; Spellerberg B; Streptococcus agalactiae is an important human pathogen causing severe neonatal infections . During the course of infection, S . agalactiae colonizes and invades a number of different host compartments . Bacterial molecules including the polysaccharide capsule, the hemolysin, the C5a peptidase, the C-proteins, the hyaluronate lyase and a number of unknown bacterial components determine the interaction with host tissues . This review summarizes our current knowledge about these interactions.

J Clin Microbiol, 2001 Jan, 39(1), 367 - 9
Use of penicillin MICs to predict in vitro activity of other beta-lactam antimicrobial agents against Streptococcus pneumoniae; Brueggemann AB et al.; Linear regression analysis was used to compare penicillin MICs determined with 3,129 recent clinical isolates of Streptococcus pneumoniae to MICs obtained with nine other beta-lactam antimicrobial agents . A strong correlation between penicillin MICs and those obtained with other beta-lactams was demonstrated . It may be possible to test penicillin and use MICs obtained with penicillin to predict MICs of other beta-lactam antimicrobials for Streptococcus pneumoniae.

Mol Microbiol, 2001 Jan, 39(2), 512 - 9
The SpeB virulence factor of Streptococcus pyogenes, a multifunctional secreted and cell surface molecule with strepadhesin, laminin-binding and cysteine protease activity; Hytonen J et al.; The interactions between pathogenic bacteria and the host need to be resolved at the molecular level in order to develop novel vaccines and drugs . We have previously identified strepadhesin, a novel glycoprotein-binding activity in Streptococcus pyogenes, which is regulated by Mga, a regulator of streptococcal virulence factors . We have now identified the protein responsible for the strepadhesin activity and find that (i) strepadhesin activity is carried by SpeB, streptococcal pyrogenic exotoxin with cysteine protease activity; (ii) SpeB carries laminin-binding activity of the bacteria; and (iii) SpeB is not only a secreted molecule but also occurs unexpectedly tightly bound to the bacterial cell surface . Thus, in contrast to the previous view of SpeB as mainly an extracellular protease, it is also present as a streptococcal surface molecule with binding activity to laminin and other glycoproteins.

Mol Microbiol, 2001 Jan, 39(2), 392 - 406
Group A streptococcal growth phase-associated virulence factor regulation by a novel operon (Fas) with homologies to two-component-type regulators requires a small RNA molecule; Kreikemeyer B et al.; A novel growth phase-associated two-component-type regulator, Fas (fibronectin/fibrinogen binding/haemolytic activity/streptokinase regulator), of Streptococcus pyogenes was identified in the M1 genome sequence, based on homologies to the histidine protein kinase (HPK) and response regulator (RR) part of the Staphylococcus aureus Agr and Streptococcus pneumoniae Com quorum-sensing systems . The fas operon, present in all 12 tested M serotypes, was transcribed as polycystronic message (fasBCA) and contained genes encoding two potential HPKs (FasB and FasC) and one RR (FasA) . Downstream of fasBCA, we identified a small 300 nucleotide monocistronic transcript, designated fasX, that did not appear to encode true peptide sequences . Measurements of luciferase promoter fusions revealed a growth phase-associated transcription of fasBCA and fasX, with peak activities during the late exponential phase . Insertional mutagenesis disrupting fasBCA and fasA led to a phenotype similar to agr-null mutations in S . aureus, with prolonged expression of extracellular matrix protein-binding adhesins and reduced expression of secreted virulence factors such as streptokinase and streptolysin S . In addition, fasX transcription was dependent on the RR FasA; however, deletion mutagenesis of fasX resulted in a similar phenotype to that of the fasBCA or fasA mutants . Complementation of the fasX deletion mutant, with the fasX gene expressed in trans from a plasmid, restored the wild-type fasBCA regulation pattern . This strongly suggested that fasX, a putative non-translated RNA, is the main effector molecule of the fas regulon . However, using spent culture supernatants from wild-type and fas mutant strains, we were not able to show an influence on the logarithmic growth phase expression of fas and dependent genes . Thus, despite structural and functional similarities between fas and agr, to date the fas operon appears not to be involved in group A streptococcal (GAS) quorum-sensing regulation.

N Engl J Med, 2000 Dec 28, 343(26), 1917 - 24
Increasing prevalence of multidrug-resistant Streptococcus pneumoniae in the United States; Whitney CG et al.; BACKGROUND: The emergence of drug-resistant strains of bacteria has complicated treatment decisions and may lead to treatment failures . METHODS: We examined data on invasive pneumococcal disease in patients identified from 1995 to 1998 in the Active Bacterial Core Surveillance program of the Centers for Disease Control and Prevention . Pneumococci that had a high level of resistance or had intermediate resistance according to the definitions of the National Committee for Clinical Laboratory Standards were defined as "resistant" for this analysis . RESULTS: During 1998, 4013 cases of invasive Streptococcus pneumoniae disease were reported (23 cases per 100,000 population); isolates were available for 3475 (87 percent) . Overall, 24 percent of isolates from 1998 were resistant to penicillin . The proportion of isolates that were resistant to penicillin was highest in Georgia (33 percent) and Tennessee (35 percent), in children under five years of age (32 percent, vs . 21 percent for persons five or more years of age), and in whites (26 percent, vs . 22 percent for blacks) . Penicillin-resistant isolates were more likely than susceptible isolates to have a high level of resistance to other antimicrobial agents . Serotypes included in the 7-valent conjugate and 23-valent pneumococcal polysaccharide vaccines accounted for 78 percent and 88 percent of penicillin-resistant strains, respectively . Between 1995 and 1998 (during which period 12,045 isolates were collected), the proportion of isolates that were resistant to three or more classes of drugs increased from 9 percent to 14 percent; there also were increases in the proportions of isolates that were resistant to penicillin (from 21 percent to 25 percent), cefotaxime (from 10 percent to 15 percent), meropenem (from 10 percent to 16 percent), erythromycin (from 11 percent to 16 percent), and trimethoprim-sulfamethoxazole (from 25 percent to 29 percent) . The increases in the frequency of resistance to other antimicrobial agents occurred exclusively among penicillin-resistant isolates . CONCLUSIONS: Multidrug-resistant pneumococci are common and are increasing . Because a limited number of serotypes account for most infections with drug-resistant strains, the new conjugate vaccines offer protection against most drug-resistant strains of S . pneumoniae.

Curr Microbiol, 2001 Feb, 42(2), 106 - 10
Regulation of H(+)-ATPase synthesis in response to reduced pH in ruminal bacteria; Miwa T et al.; The capacity of ruminal bacteria to regulate H(+)-ATPase synthesis in response to reduced pH was investigated to explain acid tolerance . The activity of H(+)-ATPase in Streptococcus bovis, an acid-tolerant bacterium, was 2.2-fold higher at pH 4.5 than at pH 5.5 . The increase in the amount of H(+)-ATPase protein was similar, suggesting that the increase in H(+)-ATPase activity is owing to the increase in H(+)-ATPase synthesis . The level of atp-mRNA at pH 4.5 was 2.5-fold higher than at pH 5.5, indicating that H(+)-ATPase synthesis is regulated at the transcriptional level, responding to low pH . In Ruminococcus albus, an acid-sensitive bacterium, H(+)-ATPase activity, the amount of H(+)-ATPase protein, and the level of atp-mRNA at pH 7.0 were similar to the values at pH 6.0, the lowest pH permitting growth . This result suggests that R . albus is incapable of enhancing H(+)-ATPase synthesis at low pH . Thus, acid tolerance appeared to be related to the capacity to augment the synthesis of H(+)-ATPase responding to low pH.

Eur J Obstet Gynecol Reprod Biol, 2001 Jan, 94(1), 79 - 85
Neonatal group B streptococcal infection . Results of 33 months of universal maternal screening and antibioprophylaxis; Volumenie JL et al.; OBJECTIVE: To assess the efficacy and pitfalls of a protocol of generalized screening for group B Streptococcus (GBS) and intra-partum treatment of all carriers in a clinical setting . DESIGN: A descriptive study and comparison with an historical group . Setting: A tertiary perinatal center . POPULATION: All women attending prenatal care in our center and delivered after 37 weeks were eligible . Study period ranged from January 1994 to September 1996 . Comparison group consisted in deliveries of years 1992 and 1993 . METHODS: Vaginal cultures were performed at 36 weeks on non-selective medium followed by intra-partum treatment of all carrier mothers . Rate of carriage, incidence of neonatal GBS sepsis, influence of risk factors and the reasons for failures were analysed . Comparison was made with an historical group . Statistical analysis was performed using a Chi-square test . RESULTS: There were 5374 term deliveries during the study . 3906 were screened (72.7%) and 559 of them found positive for GBS (14.3%) . We observed 46 early-onset GBS diseases (0.86% of term-births) . 43.5% of infections occurred in babies born from mothers without risks factors at delivery . Negative GBS cultures at sampling accounted for 43.5% of protocol failures . Comparison of the incidence of early-onset GBS disease with the previous two years showed a significant drop (1.45-0.86%, P<0.05) . CONCLUSIONS: Our protocol revealed feasible and effective in reducing the incidence of early-onset GBS disease . Improvements must be studied particularly as to the predictive value of screening cultures.

J Bacteriol, 2001 Jan, 183(2), 768 - 72
Competence modulation by the NADH oxidase of Streptococcus pneumoniae involves signal transduction; Echenique JR et al.; Oxygen controls competence development in Streptococcus pneumoniae . Oxygen signaling involves the two-component signal transduction systems CiaRH and ComDE and the competence-stimulating peptide encoded by comC and processed by ComAB . We found that NADH oxidase (Nox) was required for optimal competence . Transcriptional analysis and genetic dissection showed that Nox was involved in post-transcriptional activation of the response regulator ComE and in the transcriptional control of ciaRH and comCDE . Thus, in S . pneumoniae, Nox, with O(2) as its secondary substrate, is part of the O(2)-signaling pathway.

Appl Environ Microbiol, 2001 Jan, 67(1), 473 - 4
Pigment production by Streptococcus agalactiae in quasi-defined media; Rosa-Fraile M et al.; A quasi-defined medium that supports the growth of Streptococcus agalactiae as pigmented colonies has been developed . The medium contains starch, a peptic digest of albumin, amino acids, nucleosides, vitamins, and salts . The presence of free cysteine, which could be replaced with other sulphur-containing compounds and to a lesser degree by reducing agents, was required for pigment formation.

Appl Environ Microbiol, 2001 Jan, 67(1), 15 - 21
The group I strain of Streptococcus mutans, UA140, produces both the lantibiotic mutacin I and a nonlantibiotic bacteriocin, mutacin IV; Qi F et al.; Strains of Streptococcus mutans produce at least three mutacins, I, II, and III . Mutacin II is a member of subgroup AII in the lantibiotic family of bacteriocins, and mutacins I and III belong to subgroup AI in the lantibiotic family . In this report, we characterize two mutacins produced by UA140, a group I strain of S . mutans . One is identical to the lantibiotic mutacin I produced by strain CH43 (F . Qi et al., Appl . Environ . Microbiol . 66:3221-3229, 2000); the other is a nonlantibiotic bacteriocin, which we named mutacin IV . Mutacin IV belongs to the two-peptide, nonlantibiotic family of bacteriocins produced by gram-positive bacteria . Peptide A, encoded by gene nlmA, is 44 amino acids (aa) in size and has a molecular mass of 4,169 Da; peptide B, encoded by nlmB, is 49 aa in size and has a molecular mass of 4,826 Da . Both peptides derive from prepeptides with glycines at positions -2 and -1 relative to the processing site . Production of mutacins I and IV by UA140 appears to be regulated by different mechanisms under different physiological conditions . The significance of producing two mutacins by one strain under different conditions and the implication of this property in terms of the ecology of S . mutans in the oral cavity are discussed.

J Dairy Sci, 2000 Dec, 83(12), 2975 - 9
Efficacies of chlorine dioxide and lodophor teat dips during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae; Boddie RL et al.; We tested two postmilking teat dips for efficacy against Staphylococcus aureus and Streptococcus agalactiae using experimental challenge procedures recommended by the National Mastitis Council . The chlorine dioxide teat dip that contained 0.7% sodium chlorite reduced the number of new intramammary infections (IMI) caused by Staph . aureus by 86.6% and reduced new IMI caused by Strep . agalactiae by 88.4% . The 0.5% iodophor teat dip reduced the number of new IMI caused by Staph . aureus by 92.9% and reduced the number of new IMI caused by Strep . agalactiae by 43.4% . Teat skin and teat end conditions were evaluated before and after the study, and no deleterious effects were noted among dipped quarters compared with undipped control quarters for either teat dip.

J Biol Chem, 2001 Apr 13, 276(15), 11844 - 51 Epub 2000 Dec 15.
Crystal structure of Streptococcus pneumoniae N-acetylglucosamine-1-phosphate uridyltransferase bound to acetyl-coenzyme A reveals a novel active site architecture; Sulzenbacher G et al.; The bifunctional bacterial enzyme N-acetyl-glucosamine-1-phosphate uridyltransferase (GlmU) catalyzes the two-step formation of UDP-GlcNAc, a fundamental precursor in bacterial cell wall biosynthesis . With the emergence of new resistance mechanisms against beta-lactam and glycopeptide antibiotics, the biosynthetic pathway of UDP-GlcNAc represents an attractive target for drug design of new antibacterial agents . The crystal structures of Streptococcus pneumoniae GlmU in unbound form, in complex with acetyl-coenzyme A (AcCoA) and in complex with both AcCoA and the end product UDP-GlcNAc, have been determined and refined to 2.3, 2.5, and 1.75 A, respectively . The S . pneumoniae GlmU molecule is organized in two separate domains connected via a long alpha-helical linker and associates as a trimer, with the 50-A-long left-handed beta-helix (LbetaH) C-terminal domains packed against each other in a parallel fashion and the C-terminal region extended far away from the LbetaH core and exchanged with the beta-helix from a neighboring subunit in the trimer . AcCoA binding induces the formation of a long and narrow tunnel, enclosed between two adjacent LbetaH domains and the interchanged C-terminal region of the third subunit, giving rise to an original active site architecture at the junction of three subunits.

J Comput Aided Mol Des, 2000 Nov, 14(8), 719 - 30
Construction of a full three-dimensional model of the transpeptidase domain of Streptococcus pneumoniae PBP2x starting from its Calpha-atom coordinates; van Hooft PA et al.; A new method is described for generating all-atom protein structures from Calpha-atom information . The method, which combines both local structural trace alignments and comparative side chain modeling with ab initio side chain modeling, makes use of both the virtual-bond and the dipole-path methods . Provided that 3D structures of structurally and functionally related proteins exist, the method presented here is highly suitable for generating all-atom coordinates of partly solved, low-resolution crystal structures . Particularly the active site region can be modeled accurately with this procedure, which enables investigation of the binding modes of different classes of ligands with molecular dynamics simulations . The method is applied to the trace of Streptococcus pneumoniae, in order to construct an all-atom structure of the transpeptidase domain . Since after generation of full coordinates of the transpeptidase domain the structure had been solved to 2.4 A resolution, new X-ray coordinates for the worst modeled loop (residues T370 to M386; 17 out of a total number of 351 residues constituting the transpeptidase domain) were incorporated, as kindly provided by Dr . Dideberg . The structure was relaxed with molecular dynamics simulations and simulated annealing methods . The RMS deviation between the 144 aligned Calpha-atoms and the corresponding ones in the originally solved 3.5 A resolution crystal structure was 0.98 . The 351 Calpha-atoms of the whole transpeptidase domain of the final model showed an RMS deviation of 1.58 . The Ramachandran plot showed that 79.3% of the residues are in the most favored regions, with only 1.0% occurring in disallowed regions . The model presented here can be used to investigate the three-dimensional influences of mutations around the active site of PBP2x.

J Chemother, 2000 Nov, 12 Suppl 5, 5 - 14
Teicoplanin Chemistry and Microbiology; Parenti F et al.; The chemistry, microbiology and mode of action of teicoplanin, as well as the mechanism, control and epidemiology of glycopeptide resistance, are discussed in detail . The antibacterial activity of teicoplanin against Gram-positive bacteria, including those expressing resistance to unrelated compounds, is similar to that of vancomycin but with increased potency, particularly against Streptococcus spp and Enterococcus spp . Some strains of coagulase-negative Staphylococcus spp, particularly S . haemolytieus, are less susceptible to teicoplanin than to vancomycin . Teicoplanin is active against vancomycin resistance caused by VanB and VanC, but is not active against VanA resistant strains . The epidemiology of GISA and VISA strains of S . aureus is, as yet, poorly understood with more work necessary to elucidate the sequence of events leading to their evolution . Despite the increasing importance of glycopeptide resistance, teicoplanin has proved its clinical worth and continues to have important potential in the treatment of life-threatening Gram-positive sepsis.

J Chemother, 2000 Oct, 12 Suppl 4, 7 - 15
Fluoroquinolones: is there a different mechanism of action and resistance against Streptococcus pneumoniae?
Harding I, Simpson I.
Starting in the 1950s, study and elucidation of the biochemical mechanisms of resistance to antibiotics led to the understanding of both the biology of bacteria and the mode of action of antibiotics . This holds true for the relationship between Streptococcus pneumoniae and the fluoroquinolones . A new feature in this approach is the availability of the nearly complete chromosome sequence of this major human pathogen . In S . pneumoniae, resistance appears to be mainly due to mutational alterations in the intracellular targets of the fluoroquinolones, the type II DNA topoisomerase gyrase and topoisomerase IV . Both enzymes appear to be the primary targets of the drugs in this species . Mutations in the quinolone resistance-determining region (QRDR) of the gyrA gene or the parC gene, which encode the A subunits of DNA gyrase and topoisomerase IV respectively, confer resistance to single-step mutants . Mutations in gyrB and parE, which encode the B subunits of DNA gyrase and topo IV, respectively, have also been implicated in the fluoroquinolone resistance of certain mutants obtained in vitro . The antibiotics most affected by a single mutation are those for which the mutation occurs in their preferred target e.g . gyrase for sparfloxacin and gatifloxacin and topo IV for ciprofloxacin and levofloxacin . The activity of all fluoroquinolones is decreased further when two or more mutations are present . Because they possess similar targets of action, there is cross resistance, albeit at various degrees depending on the intrinsic activity of the molecule, among fluoroquinolones . This stresses, once more, the misleading concept of breakpoints for clinical categorization . A second mechanism of resistance, enhanced active efflux of hydrophilic quinolones such as norfioxacin and ciprofloxacin, is mediated by the membrane-associated protein, PmrA (pneumococcal multidrug resistance) . This protein is a 12-transmembrane segment proton-dependent multidrug efflux pump of the major facilitator family . The combinatorial approach of bacteria to fluoroquinolone resistance implies that the molecule actually used, as well as a less active member of the class that is more apt to detect resistance mechanisms (e.g . ciprofloxacin), should be tested in vitro.

J Chemother, 2000 Oct, 12 Suppl 4, 27 - 31
The use of levofloxacin in the treatment of respiratory tract infection; Shah PM; Increasing resistance among the common respiratory pathogens has encouraged assessment of alternative agents, for example, levofloxacin . Unlike earlier quinolones, levofloxacin has excellent activity against Streptococcus pneumoniae, including strains resistant to penicillin . Clinical trials show levofloxacin to be as effective as cephalosporins in acute exacerbation of chronic bronchitis and as effective as co-amoxiclav, cephalosporins or amoxycillin in community-acquired pneumonia . Levofloxacin is rarely associated with serious adverse events . Nausea, diarrhea, headache and rash are the most common adverse events but are observed less frequently than with some other new quinolones.

Am J Vet Res, 2000 Dec, 61(12), 1525 - 9
Pathogenesis of Streptococcus zooepidemicus infection after intratracheal inoculation in llamas; Cebra CK et al.; OBJECTIVES: To test whether generalized Streptococcus zooepidemicus infection could be induced by intratracheal inoculation in llamas and to characterize this infection . ANIMALS: 6 test and 3 control llamas . PROCEDURE: Test llamas received 1 of 3 dosages of S . zooepidemicus by intratracheal injection, whereas control llamas received sterile culture medium . Physical examination variables and results of clinicopathologic analyses of blood, peritoneal fluid, and tracheal wash fluid were compared in test llamas between, before, and during the development of bacteremia and with control llamas . Bacteriologic culture was performed on all collected body fluids and tissue specimens that were collected at necropsy . Tissue specimens that were collected at necropsy were examined histologically . RESULTS: Infection induced fever, anorexia, and signs of depression . Five of 6 infected llamas developed specific signs of inflammation in the thorax or abdomen, bacteremia, neutrophilic leukocytosis with toxic changes and high band neutrophil cell counts, hyperfibrinogenemia, and high peritoneal fluid WBC counts and protein concentrations . On development of bacteremia, llamas had significant decreases in serum iron (from 118+/-25 to 6+/-4 microg/ml) and increases in serum glucose (from 131+/-5 to 253+/-48 mg/dl) concentrations . CONCLUSIONS AND CLINICAL RELEVANCE: Streptococcus zooepidemicus spreads rapidly to other body compartments after intratracheal inoculation in llamas . Fever, anorexia, and signs of depression are the most consistent clinical signs, although other signs are possible . Clinicopathologic analysis of body fluids yields evidence of inflammation . Infection by S . zooepidemicus can be proven by bacteriologic culture of body fluids before death or of tissue specimens after death.

Scott Med J, 2000 Oct, 45(5), 153 - 4
Empyema of lung associated with Streptococcus milleri infection; Palaniappan S et al.; Empyema of the lung is a very serious illness which must be detected quickly and treated aggressively . We report an unusual case of empyema of the lung associated with a boating accident while the patient was fishing in a sea loch off the west coast of Scotland.

J Am Coll Surg, 2000 Dec, 191(6), 668 - 71
Evaluation of phagocytic function of macrophages in rats after partial splenectomy; Muftuoglu TM et al.; BACKGROUND: Understanding the immunologic properties of the spleen has enabled surgeons to practice splenic conservation surgery . If the upper pole of the spleen can be preserved solely on the upper short gastric vessels, will phagocytic function of macrophages in remnant splenic tissue be affected? The aim of this experimental study was to evaluate the phagocytic function of macrophages in partially resected spleens, with hilar excision preserving the short gastric vessels . STUDY DESIGN: Forty-eight female Wistar albino rats were divided into four groups . Groups 1 and 2 underwent sham operations and groups 3 and 4 underwent partial splenectomy . One milliliter of sodium chloride 0.9% was injected into the abdomen of the rats in groups 1 and 3 and 1 mL of Streptococcus pneumoniae type III as an antigenic stimulus was injected into the abdomen of the rats in groups 2 and 4, 6 weeks after the first operation . Forty-eight hours later, relaparotomy was performed in all animals . India ink was used to determine the capacity of uptake in the splenic phagocytes . To evaluate the phagocytic function of the splenic tissues, histologic examinations were performed according to a macrophage grading system . RESULTS: All spleens in all four groups were stained black after injection of India ink . Phagocytic activity of macrophages was reduced in the partially splenectomized groups, compared with intact spleen groups (group 3 versus group 1; p < 0.0001, group 4 versus group 2; p < 0.0001) . There was a significant difference between groups 1 and 2 according to phagocytic function of macrophages (p = 0.0121) . Also, after Streptococcus pneumoniae type III injection as an antigenic stimulus in group 4, we found that the phagocytic functions of macrophages increased compared with those of the sodium chloride 0.9%-injected group 3 after partial splenectomy (p < 0.0001) . CONCLUSIONS: Phagocytic function of macrophages in rats decreased after partial splenectomy . Nevertheless, the remnant spleens in rats could be stimulated when challenged with an antigenic stimulus.

J Chemother, 2000 Oct, 12(5), 406 - 11
Moraxella catarrhalis pneumonia during HIV disease; Manfredi R et al.; To assess the role of Moraxella catarrhalis complications in the setting of HIV disease, and to evaluate their occurrence and outcome according to several epidemiological, clinical, and laboratory parameters, the clinical records of 2123 consecutive HIV-infected patients hospitalized in a 9-year period were retrospectively reviewed, and 4 cases of community-acquired M . catarrhalis pneumonia were identified . Three adult patients had a diagnosis of AIDS and severe concurrent immunodeficiency (with a CD4+ lymphocyte count below 60 cells/microL), while the fourth case involved a child with vertical HIV disease . Leukopenia and neutropenia were never present, but no patient received a potent antiretroviral regimen at the time of disease onset . A concurrent respiratory infection by Streptococcus pneumoniae and Mycobacterium tuberculosis was recognized in 2 of 4 patients . Isolated M . catarrhalis strains were susceptible to all tested antimicrobial compounds (save ampicillin in 2 cases), and appropriate antimicrobial treatment led to clinical and microbiological cure in all described episodes . Only 8 cases of HIV-associated Moraxella spp . disease have been reported to date in seven different literature reports (6 cases of pneumonia, and 1 of septicemia) . According to our experience, M . catarrhalis may be responsible for appreciable morbidity among patients with advanced HIV infection, especially when a low CD4+ cell count or coexisting respiratory disease are present . Clinicians and microbiologists who care for HIV-infected patients should carefully consider the potential pathogenic role of Moraxella spp . organisms.

Microbiol Immunol, 2000, 44(10), 863 - 5
Erythromycin resistance genes in Streptococcus pyogenes isolates in Kanagawa, Japan; Murase T et al.; The susceptibility of 224 Streptococcus pyogenes isolates obtained from children in Japan from 1981 to 1997 to treatment with erythromycin was determined by the agar dilution method . A total of 17 isolates belonging to serotype M12T12 were resistant (MICs>1 microg/ml) . Fourteen of the 17 resistant strains obtained from 1982 to 1985 harbored ermB and showed an identical pulsed-field gel electrophoresis pattern, indicating the spread of a single clone . Two ermTR-containing isolates were obtained in 1983 . mefA gene was found in a strain obtained in 1994 in the present study, although this gene is predominantly associated with recent erythromycin resistance among S . pyogenes strains in many countries.

Clin Orthop, 2000 Dec, (381), 101 - 5
Limited role of direct exchange arthroplasty in the treatment of infected total hip replacements; Jackson WO et al.; A literature review was performed to determine when direct exchange was most likely to be successful . Twelve reports provided outcome data on infected hip replacements treated with direct exchange . The average duration of followup was 4.8 years, but the range was broad (0.1-17.1 years) . Of the 1,299 infected hip replacements treated with direct exchange, 1,077 (83%) were thought to be free of infection at the last followup . Antibiotic-impregnated bone cement was used in 1,282 of the cases (99%) . There was wide variability in the duration of parenteral antibiotic therapy, ranging from just 24 hours to as many as 8 weeks . In some cases, no oral antibiotics ever were given, whereas in others, oral antibiotics were given for as many as 8 months after parenteral therapy . Factors associated with a successful direct exchange included: (1) absence of wound complications after the initial total hip replacement; (2) good general health of the patient; (3) methicillin-sensitive Staphylococcus epidermidis, Staphylococcus aureus, and Streptococcus species; and (4) an organism that was sensitive to the antibiotic mixed into the bone cement . Factors associated with failure included: (1) polymicrobial infection; (2) gram-negative organisms, especially Pseudomonas species; and (3) certain gram-positive organisms such as methicillin-resistant Staphylococcus epidermidis and Group D Streptococcus . Methicillin-resistant organisms have become more common . Many current revision surgical techniques use cementless implants . Fixation without any cement (no depot antibiotics) may be a contraindication to direct exchange . Additionally, there essentially are no data on the use of bone graft in association with direct exchange . For these reasons, the indications for direct exchange are limited.

Salud Publica Mex, 2000 Sep-Oct, 42(5), 413 - 21
{Factors associated with Streptococcus group B colonization in pregnant women in Los Altos, Chiapas}; Ocampo-Torres M et al.; OBJECTIVE: To estimate the prevalence and analyze the factors associated with group B Streptococcus (GBS) colonization in pregnant women of Los Altos, Chiapas, Mexico . MATERIAL AND METHODS: Between February and September 1999, a cross-sectional study was conducted among 910 women who sought delivery care at three public hospitals of San Cristobal de Las Casas, Chiapas . Vaginal and perianal samples were taken for GBS detection by bacteriological culture . Identification of groups and serotypes was performed using latex agglutination . The analysis of factors associated with colonization was done using chi-squared tests and log-linear modeling . RESULTS: GBS colonization was found in 8.6% (95% CI 6.8-10.5) of study subjects . Women with the greatest likelihood of colonization were those with > or = 5 pregnancies, residents of counties with high levels of poverty, working outside the home, and living in homes in which the head of household worked in agriculture (26.8%, OR = 7.25, 95% CI 1.83-28.67) . CONCLUSIONS: In the study area, it is necessary that actions aiming to prevent and control infections by GBS be directed principally at those groups of women with the highest probability of colonization, in order to diminish the perinatal transmission of GBS.

Chemotherapy, 2001 Jan-Feb, 47(1), 39 - 42
In vitro development of resistance to three quinolones in Streptococcus pneumoniae; Rodriguez JC et al.; We studied the in vitro development of resistance to ciprofloxacin, trovafloxacin and moxifloxacin in 5 strains of Streptococcus pneumoniae resistant to penicillin . We detected the great ease of in vitro development of resistance in the case of ciprofloxacin and the much reduced capacity of moxifloxacin to generate resistance (only 1 strain) . Trovafloxacin generated resistance, but more slowly than ciprofloxacin . We consider that study of the capacity to generate resistance should be one of the points to consider when deciding on their large-scale use in respiratory infections, but comparative studies between in vitro and in vivo models should be carried out so as to determine the clinical repercussion of these phenomena .

Caries Res, 2001 Jan-Feb, 35(1), 67 - 74
Properties of Streptococcus sanguinis glucans formed under various conditions; Kopec LK et al.; The aim of our study was to determine whether the structure of glucans formed by glucosyltransferase from Streptococcus sanguinis (GtfSs) on a surface differ from those formed in solution and to explore the effects of antiserum to Gtfs, control normal rabbit serum, starch hydrolysates (STH) and dextran on S . sanguinis (GtfSs) glucan . Linkage analyses showed that solution-formed glucans are predominantly alpha-1,6-linked and have a small amount of alpha-1,3-linked glucose . Surface-formed glucans have enhanced susceptibility to mutanase . Solution- and surface-formed glucans made in the presence or absence of sera, STH, and dextran contain linkages which differ in both amount and type from control glucans . The GtfSs enzyme in solution exposed to antiserum behaves as if it is adsorbed to a surface . Binding of Streptococcus mutans GS-5 and Actinomyces viscosus OMZ105E (Ny1) to S . sanguinis glucan differs if the glucan is formed in the presence of antiserum . The information could help to define the role of glucans in the formation of pellicle, colonization of tooth surfaces and the accumulation of dental plaque.

J Mol Biol, 2001 Jan 12, 305(2), 279 - 89
Crystal structures of Streptococcus pneumoniae N-acetylglucosamine-1-phosphate uridyltransferase, GlmU, in apo form at 2.33 A resolution and in complex with UDP-N-acetylglucosamine and Mg(2+) at 1.96 A resolution; Kostrewa D et al.; N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) is an essential bacterial enzyme with both an acetyltransferase and a uridyltransferase activity which have been mapped to the C-terminal and N-terminal domains, respectively . GlmU performs the last two steps in the synthesis of UDP-N-acetylglucosamine (UDP-GlcNAc), which is an essential precursor in both the peptidoglycan and the lipopolysaccharide metabolic pathways . GlmU is therefore an attractive target for potential antibiotics . Knowledge of its three-dimensional structure would provide a basis for rational drug design . We have determined the crystal structures of Streptococcus pneumoniae GlmU (SpGlmU) in apo form at 2.33 A resolution, and in complex with UDP-N-acetyl glucosamine and the essential co-factor Mg(2+) at 1.96 A resolution . The protein structure consists of an N-terminal domain with an alpha/beta-fold, containing the uridyltransferase active site, and a C-terminal domain with a long left-handed beta-sheet helix (LbetaH) domain . An insertion loop containing the highly conserved sequence motif Asn-Tyr-Asp-Gly protrudes from the left-handed beta-sheet helix domain . In the crystal, S . pneumoniae GlmU forms exact trimers, mainly through contacts between left-handed beta-sheet helix domains . UDP-N-acetylglucosamine and Mg(2+) are bound at the uridyltransferase active site, which is in a closed form . We propose a uridyltransferase mechanism in which the activation energy of the double negatively charged phosphorane transition state is lowered by charge compensation of Mg(2+) and the side-chain of Lys22 .

Mol Microbiol, 2001 Jan, 39(1), 126 - 35
Differential fluorescence induction reveals Streptococcus pneumoniae loci regulated by competence stimulatory peptide; Bartilson M et al.; Differential fluorescence induction (DFI) in Streptococcus pneumoniae was used as a method for the discovery of genes activated in specific growth environments . Competence stimulatory peptide (CSP) was used as the model inducing system to identify differentially expressed genes . To identify CSP-induced promoters, a plasmid library was constructed by inserting random pieces of S . pneumoniae chromosomal DNA upstream of the promoterless gfpmut2 gene in an Escherichia coli/S . pneumoniae shuttle vector . S . pneumoniae carrying the library were induced with CSP and enriched for green fluorescent protein (GFP)-expressing bacteria using fluorescence-activated cell sorting . A total of 886 fluorescent clones was screened, and 12 differentially activated promoter elements were identified . Sequence analysis of these clones revealed that three were associated with novel competence loci, one of which we show is essential for DNA uptake, and six are known CSP-inducible promoters . We also explored whether competence proteins have a role in virulence and found that mutations in three CSP-inducible genes resulted in attenuated virulence phenotypes in either of two murine infection models . These results demonstrate the utility of DFI as a method for identifying differentially expressed genes in S . pneumoniae and the potential utility of applying DFI to other Gram-positive bacteria.

J Struct Biol, 2000 Oct, 132(1), 72 - 81
Structure and molecular mechanism of a functional form of pneumolysin: a cholesterol-dependent cytolysin from Streptococcus pneumoniae; Kelly SJ et al.; One of the key steps in understanding human disease arising from gram-positive bacteria lies in the mechanisms of the cholesterol-dependent cytolysins (CDCs) . Pneumolysin (PLY), a CDC from Streptococcus pneumoniae, is of special importance due to the severe impacts of pneumococcal infections on mortality and morbidity worldwide . We have overexpressed, purified, and characterized PLY in its fully functional complex form with the enzyme bound to its receptor activator on target cells, cholesterol . The circular dichroism studies of PLY in solution with an excess of cholesterol show a change in the far UV spectrum consistent with a decrease in the beta-sheet and an increase in the random coil structures of the enzyme . Pore formation in membranes leading to cell lysis is the functional target for this cytolysin . The sedimentation velocity and equilibrium analyses of the cholesterol-bound enzyme show hydrodynamic properties different from those of the cholesterol-free form . The soluble form of the cholesterol-free enzyme exists in solution as a mixture of monomers and dimers, whereas the cholesterol-bound form exists only as a monomer . A mechanism of formation of PLY pores in the lipid bilayer of the target cells is discussed .

Antimicrob Agents Chemother, 2001 Jan, 45(1), 342 - 4
Identification of an erm(A) erythromycin resistance methylase gene in Streptococcus pneumoniae isolated in Greece; Syrogiannopoulos GA et al.; In a serotype 11A clone of erythromycin-resistant pneumococci isolated from young Greek carriers, we identified the nucleotide sequence of erm(A), a methylase gene previously described as erm(TR) in Streptococcus pyogenes . The erm(A) pneumococci were resistant to 14- and 15-member macrolides, inducibly resistant to clindamycin, and susceptible to streptogramin B . To our knowledge, this is the first identification of resistance to erythromycin in S . pneumoniae attributed solely to the carriage of the erm(A) gene.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 339 - 41
Prevalence and mechanisms of macrolide resistance in Streptococcus pyogenes in Santiago, Chile; Palavecino EL et al.; Thirty-two macrolide-resistant Streptococcus pyogenes isolates were found among 594 clinical isolates collected from 1990 to 1998 in Santiago, Chile, for an overall prevalence of 7.2% . Among the 32 resistant isolates, 28 (87.5%) presented the M phenotype and 4 (12 . 5%) presented the MLS(B) phenotype . Serotyping and pulsed-field gel electrophoresis analysis showed genetic diversity among the resistant isolates.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 319 - 23
Mutation in 23S rRNA responsible for resistance to 16-membered macrolides and streptogramins in Streptococcus pneumoniae; Depardieu F et al.; Streptococcus pneumoniae clinical isolate BM4455 was resistant to 16-membered macrolides and to streptogramins . This unusual resistance phenotype was due to an A(2062)C (Escherichia coli numbering) mutation in domain V of the four copies of 23S rRNA.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 316 - 8
Effective combination therapy for invasive pneumococcal pneumonia with ampicillin and intravenous immunoglobulins in a mouse model; De Hennezel L et al.; Intranasal immunotherapy for Streptococcus pneumoniae invasive pneumonia with polyvalent immunoglobulins (IVIG) was effective in mice against pneumonia but failed to prevent bacteremia . The combination of subcurative doses of IVIG and of ampicillin was fully protective . Such an approach, successfully applied in the preantibiotic era, offers new perspectives for modern therapies.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 252 - 62
Kinetic study of the inflammatory response in Streptococcus pneumoniae experimental pneumonia treated with the ketolide HMR 3004; Duong M et al.; Patients still die from Streptococcus pneumoniae pneumonia after initiation of antibiotic therapy, when tissues are sterile and the pneumonia is clearing . There is growing evidence that overwhelming inflammation resulting from toxin release contributes to tissue injury, shock, and death . Monitoring host response may help us understand the consequences of antibiotic therapy for the inflammatory processes that occur in bacterial pneumonia . HMR 3004 is a ketolide that displays excellent in vitro activity against S . pneumoniae . In the present experiment, we investigated the chronology of inflammatory events that occur during pneumococcal pneumonia in mice treated with HMR 3004 . Infection of mice with 10(7) CFU of living S . pneumoniae resulted in 100% mortality within 5 days . HMR 3004 given at 12.5 mg/kg of body weight/dose twice daily from 48 h postinfection achieved complete bacterial clearance from lungs and blood within 36 h and ensured survival of mice . Recruitment of neutrophils and monocytes from blood to lungs was significantly reduced, and nitric oxide release was totally prevented . Interleukin-6 secretion in lungs and blood became rapidly undetectable after initiation of therapy . Histological examination of lung tissue showed protection of interstitium against edema . By controlling bacterial invasion, HMR 3004 led to rapid and profound modifications of the host response in lungs, which may protect mice from deleterious inflammatory reactions.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 166 - 9
Effect of xylitol on growth of Streptococcus pneumoniae in the presence of fructose and sorbitol; Tapiainen T et al.; Xylitol is effective in preventing acute otitis media by inhibiting the growth of Streptococcus pneumoniae . To clarify this inhibition we used fructose, which is known to block similar growth inhibition observed in Streptococcus mutans . In addition, we evaluated the efficacy of sorbitol in inhibiting the growth of pneumococci, as sorbitol is widely used for indications similar to those for which xylitol is used . The addition of 5% xylitol to the growth medium resulted in marked growth inhibition, an effect which was totally eliminated in the presence of 1, 2.5, or 5% fructose but not in the presence of 1 or 5% glucose, 1% galactose, or 1% sucrose . This finding implies that xylitol-induced inhibition of pneumococcal growth is mediated via the fructose phosphotransferase system in a way similar to that in which mutans group streptococcal growth is inhibited . The addition of sorbitol at concentrations of 1, 2.5, or 5% to the growth medium did not affect the growth of pneumococci and neither inhibited nor enhanced the xylitol-induced growth impairment . Thus, it seems that xylitol is the only commercially used sugar substitute proven to have an antimicrobial effect on pneumococci.

J Immunol, 2000 Dec 15, 165(12), 6840 - 8
B7 requirements for primary and secondary protein- and polysaccharide-specific Ig isotype responses to Streptococcus pneumoniae; Wu ZQ et al.; The requirements for B7 costimulation during an in vivo humoral response to an intact extracellular bacteria have not been reported . In this study we immunized mice with Streptococcus pneumoniae (R36A) to determine the B7 requirements for induction of Ig, specific for two determinants on R36A, the phosphorylcholine (PC) determinant of C-polysaccharide and pneumococcal surface protein A (PspA) . We show that the primary anti-PspA response, the development of PspA-specific memory, and the induction of the secondary anti-PspA response in primed mice were completely dependent upon B7 costimulation . Of note, costimulation was required only briefly after the secondary immunization compared with after the primary immunization for optimal induction of Ig . Blockade of B7 costimulation at the time of secondary immunization also completely abrogated the established state of memory, but did not induce tolerance . In contrast to the anti-PspA response, the primary anti-PC response involved only a very short period of B7 costimulation . Whereas B7-2 alone was required for induction of the primary anti-PspA and anti-PC responses, a redundant role for B7-1 and B7-2 was noted for the PspA-specific secondary response . CTLA4Ig blocked both the anti-PC and anti-PspA responses equally well over a wide range of bacterial doses . These studies demonstrate a critical, but variable, role for B7-dependent costimulation during an Ig response to an extracellular bacteria.

Infect Immun, 2001 Jan, 69(1), 622 - 5
Characterization of the domain of fibronectin-binding protein I of Streptococcus pyogenes responsible for elicitation of a protective immune response; Schulze K et al.; Fibronectin-binding protein I (SfbI) represents a major adhesin of Streptococcus pyogenes . Mice were intranasally immunized with recombinant proteins spanning different portions of SfbI to identify the minimal fragment able to elicit a protective response against a lethal challenge with S . pyogenes . The strongest cellular responses and the highest levels of antigen-specific secretory immunoglobulin A (IgA) were detected in mice immunized with the fibronectin-binding region of SfbI . In contrast, animals vaccinated with a polypeptide spanning the aromatic and proline-rich regions showed the highest titers and fastest IgG response in serum . Vaccination with either SfbI without a membrane anchor and signal peptide or a polypeptide encompassing its fibronectin-binding regions resulted in efficient protection against heterologous challenge (60% and 80%, respectively), whereas the use of a polypeptide lacking this region conferred marginal protection (10%) with respect to the control group (0%) . These results demonstrate that the fibronectin-binding region of SfbI is a promising candidate antigen for developing anti-S . pyogenes vaccines.

Infect Immun, 2001 Jan, 69(1), 602 - 6
Effect of influenza A virus infection on nasopharyngeal colonization and otitis media induced by transparent or opaque phenotype variants of Streptococcus pneumoniae in the chinchilla model; Tong HH et al.; Phase variation in the colonial opacity of Streptococcus pneumoniae has been implicated as a factor in bacterial adherence, colonization, and invasion in the pathogenesis of pneumococcal disease . Additionally, the synergistic effects of influenza A virus and S . pneumoniae in the development of otitis media (OM) have been reported . This study examined the ability of opaque or transparent S . pneumoniae from the same strain in combination with an antecedent influenza A virus infection to colonize the nasopharynx and invade the middle ear in the chinchilla model . Our data indicated that there was no significant difference in the level of nasopharyngeal colonization and induction of OM between the opaque and transparent variants unless there was a prior challenge with influenza A virus . Subsequent to influenza A virus infection, there was a significant difference between the variants in the ability to colonize and persist in the nasopharynx and middle ear . The concentrations of the opaque variant in nasopharyngeal-lavage samples and middle-ear fluid remained consistently higher than those of the transparent variant for 10 days postinoculation . Data from this study indicate that the effects of influenza A virus on the pathogenesis of experimental S . pneumoniae-induced OM differ depending on the opacity phenotype involved.

Infect Immun, 2001 Jan, 69(1), 426 - 34
Role of genetic resistance in invasive pneumococcal infection: identification and study of susceptibility and resistance in inbred mouse strains; Gingles NA et al.; From a panel of nine inbred mice strains intranasally infected with Streptococcus pneumoniae type 2 strain, BALB/c mice were resistant and CBA/Ca and SJL mice were susceptible to infection . Further investigation revealed that BALB/c mice were able to prevent proliferation of pneumococci in the lungs and blood, whereas CBA/Ca mice showed no bacterial clearance . Rapidly increasing numbers of bacteria in the blood was a feature of CBA/Ca but not BALB/c mice . In the lungs, BALB/c mice recruited significantly more neutrophils than CBA/Ca mice at 12 and 24 h postinfection . Inflammatory lesions in BALB/c mice were visible much earlier than in CBA/Ca mice, and there was a greater cellular infiltration into the lung tissue of BALB/c mice at the earlier time points . Our data suggest that resistance or susceptibility to intranasal pneumococci may have an association with recruitment and/or function of neutrophils.

Infect Immun, 2001 Jan, 69(1), 392 - 9
Identification and disruption of two discrete loci encoding hyaluronic acid capsule biosynthesis genes hasA, hasB, and hasC in Streptococcus uberis; Ward PN et al.; The hyaluronic acid capsule of Streptococcus uberis has been implicated in conferring resistance to phagocytosis by bovine neutrophils . Construction of a bank of random insertion mutants of S . uberis (strain 0140J) was achieved using the pGh9::ISS1 mutagenesis system (22) . Phenotypic screening of approximately 5,000 clones enabled the isolation of 11 acapsular mutants . Southern hybridization indicated that two mutants carried a lesion within a group of genes similar to those involved in the assembly of the hyaluronic acid capsule found in the group A Streptococcus (GAS) has operon . The DNA sequence flanking the points of insertion confirmed the presence of homologues of GAS hasA and hasB in S . uberis . The DNA sequence flanking the ISS1 insertion in another mutant identified a homologue of hasC in S . uberis . The GAS hasABC operon structure was not conserved in S . uberis, and two discrete loci comprising homologues of either hasAB or hasC were identified . Disruption of S . uberis hasA or hasC resulted in the complete cessation of hyaluronic acid capsule production . Correspondingly, these mutants were found to have lost their resistance to phagocytosis by bovine neutrophils . The bactericidal action of bovine neutrophils on S . uberis 0140J was shown unequivocally to depend upon the capsule status of the bacterium.

Infect Immun, 2001 Jan, 69(1), 336 - 44
Avidity, potency, and cross-reactivity of monoclonal antibodies to pneumococcal capsular polysaccharide serotype 6B; Sun Y et al.; Many pneumococcal capsular polysaccharides (PSs) are similar in structure, and a pneumococcal antibody often binds to all of the PSs with a similar structure . Yet, these cross-reactive antibodies may bind to the structurally related pneumococcal capsular PSs with an avidity too low to be effective . If memory B cells producing such weakly cross-reactive antibodies are elicited with pneumococcal conjugate vaccines, the memory cells for low-avidity antibodies could compromise the subsequent immune responses to the cross-reactive PS (original antigenic sin) . To investigate these issues, we produced 14 hybridomas secreting monoclonal antibodies (MAbs) to the capsular PS of Streptococcus pneumoniae serotype 6B by immunizing BALB/c mice with antigens containing 6B PS and studied their epitope, avidity, in vitro opsonizing capacity, in vivo protective capacity, and "antigen binding titer" by enzyme-linked immunosorbent assay (ELISA) of 6A and 6B capsular PSs . Six MAbs bound to the non-cross-reactive 6B-specific epitope, and seven MAbs bound to the cross-reactive epitope present in both 6A and 6B PSs One MAb (Hyp6BM6) revealed a novel epitope . This epitope was found on 6A PS in solution, but not on 6A PS adsorbed onto the plastic surface of the ELISA plates . The avidity of the MAb for 6A or 6B PS ranged from 7.8 x 10(6) M(-1) to 4.1 x 10(11) M(-1) . No MAbs were weakly cross-reactive, since none of the cross-reactive MAbs showed any tendency toward having less avidity to 6A PS (the cross-reactive PS) than to 6B PS . Avidity influenced the results of several antibody assays . When all of the hybridomas were examined, avidity strongly correlated with the titer of a unit amount of MAb to bind antigen-coated ELISA plates (r = 0.91) or to opsonize pneumococci in vitro (r = -0.85) . Because both assay results are avidity dependent, the ELISA and the opsonization assay results were strongly correlated (r = 0.91), regardless of avidity . Avidity also correlated with the potency of a MAb to passively protect mice against pneumococcal infections . When only the immunoglobulin G hybridomas were examined, little increase in opsonizing capacity and in vivo protective potency was observed above 10(9) M(-1) . Taken together, an ELISA measuring antigen binding titer may be an adequate measure of the protective immunity induced with pneumococcal vaccines, and the absence of a partially cross-reactive MAb suggests that antigenic sin may not be significant in responses to vaccines against the S . pneumoniae 6B serotype.

Infect Immun, 2001 Jan, 69(1), 297 - 306
Group B Streptococcus capsular polysaccharide-cholera toxin B subunit conjugate vaccines prepared by different methods for intranasal immunization; Shen X et al.; Group B Streptococcus (GBS) type III capsular polysaccharide (CPS III) was conjugated to recombinant cholera toxin B subunit (rCTB) using three different methods which employed (i) cystamine and N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), (ii) carbodiimide with adipic acid dihydrazide (ADH) as a spacer, or (iii) reductive amination (RA) . The CPS III-rCTB conjugates were divided into large- and small-molecular-weight (M(r)) fractions, and the immunogenicities of the different preparations after intranasal (i.n.) immunization were studied in mice . Both large- and small-M(r) conjugates of CPS III-rCTB(RA) or CPS III-rCTB(ADH) induced high, almost comparable levels of CPS-specific immunoglobulin G (IgG) in serum, lungs, and vagina that were generally superior to those obtained with CPS III-rCTB(SPDP) conjugates or a CPS III and rCTB mixture . However, the smaller-M(r) conjugates of CPS III-rCTB(RA) or CPS III-rCTB(ADH) in most cases elicited a lower anti-CPS IgA immune response than the large-M(r) conjugates, and the highest anti-CPS IgA titers in both tissues and serum were obtained with the large-M(r) CPS III-rCTB(RA) conjugate . Serum IgG anti-CPS titers induced by the CPS III-rCTB(RA) conjugate had high levels of specific IgG1, IgG2a, IgG2b, and IgG3 antibodies . Based on the effectiveness of RA for coupling CPS III to rCTB, RA was also tested for conjugating GBS CPS Ia with rCTB . As for the CPS III-rCTB conjugates, the immunogenicity of CPS Ia was greatly increased by conjugation to rCTB . Intranasal immunization with a combination of CPS Ia-rCTB and CPS III-rCTB conjugates was shown to induce anti-CPS Ia and III immune responses in serum and lungs that were fully comparable with the responses to immunization with the monovalent CPS Ia-rCTB or CPS III-rCTB conjugates . These results suggest that the GBS CPS III-rCTB and CPS Ia-rCTB conjugates prepared by the RA method may be used in bivalent and possibly also in multivalent mucosal GBS conjugate vaccines.

Infect Immun, 2001 Jan, 69(1), 75 - 80
Inactivation of the srtA gene in Streptococcus gordonii inhibits cell wall anchoring of surface proteins and decreases in vitro and in vivo adhesion; Bolken TC et al.; The srtA gene product, SrtA, has been shown to be required for cell wall anchoring of protein A as well as virulence in the pathogenic bacterium Staphylococcus aureus . There are five major mechanisms for displaying proteins at the surface of gram-positive bacteria (P . Cossart and R . Jonquieres, Proc . Natl . Acad . Sci . USA 97:5013-5015, 2000) . However, since many of the known surface proteins of gram-positive bacteria are believed to be exported and anchored via the sortase pathway, it was of interest to determine if srtA plays a similar role in other gram-positive bacteria . To that end, the srtA gene in the human oral commensal organism Streptococcus gordonii was insertionally inactivated . The srtA mutant S . gordonii exhibited a marked reduction in quantity of a specific anchored surface protein . Furthermore, the srtA mutant had reduced binding to immobilized human fibronectin and had a decreased ability to colonize the oral mucosa of mice . Taken together, these results suggest that the activity of SrtA plays an important role in the biology of nonpathogenic as well as pathogenic gram-positive cocci.

Vet Microbiol, 2001 Jan 5, 78(1), 29 - 37
Influence of ampicillin, ceftiofur, attenuated live PRRSV vaccine, and reduced dose Streptococcus suis exposure on disease associated with PRRSV and S . suis coinfection; Schmitt CS et al.; The objective of this research was to evaluate the efficacy of two antimicrobials (ampicillin and ceftiofur), a modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine, and low dose exposure to Streptococcus suis on disease associated with PRRSV/S . suis coinfection . Fifty-six, crossbred, PRRSV-free pigs were weaned at 10-12 days of age and randomly assigned to five treatment groups . All pigs were inoculated with 2ml of 10(6.4) TCID50/ml of high virulence PRRSV isolate VR-2385 intranasally at 29-31 days of age (day 0 of the study) followed 7 days later by intranasal inoculation with 2ml of 10(8.9)colony forming units(CFU)/ml S . suis type 2 isolate ISU VDL #40634/94 . Pigs in group 1 (n=10) served as untreated infected positive controls . Pigs in group 2 (n=12) were treated with 5.0 mg/kg ceftiofur hydrochloride intramuscularly (IM) on days 8, 11, and 14 . Pigs in group 3 (n=11) were treated with 11 mg/kg ampicillin IM on days 8-10 . Pigs in group 4 (n=12) were vaccinated 14 days prior to PRRSV challenge with a commercial modified-live PRRSV vaccine . Pigs in group 5 (n=11) were exposed to a 1:100 dilution of the S . suis challenge inoculum 19 days prior to S . suis challenge . Mortality was 80, 25, 82, 83, and 36% in groups 1-5, respectively . The reduced dose S . suis exposure had some residual virulence, evidenced by S . suis induced meningitis in two pigs after exposure . Treatment with ceftiofur hydrochloride and reduced dose exposure to S . suis were the only treatments which significantly (P<0.05) reduced mortality associated with PRRSV/S . suis coinfection, significantly (P<0.05) reduced recovery of S . suis from tissues at necropsy, and significantly (P<0.05) reduced the severity of gross lung lesions.

Toxicology, 2000 Nov 23, 154(1-3), 85 - 101
Carbon tetrachloride is immunosuppressive and decreases host resistance to Listeria monocytogenes and Streptococcus pneumoniae in female B6C3F1 mice; Guo TL et al.; Carbon tetrachloride (CCl(4)) is an environmental contaminant that has been detected in ambient air, seawater, surface-water and snow . The immunotoxic potential of CCl(4) was evaluated in female B6C3F1 mice . The animals were administered with CCl(4) daily for 14 days at doses of 50, 100, 500 or 1000 mg/kg body weight by gavage with corn oil as a vehicle . Exposure to CCl(4) resulted in an increase of liver weight but not the body weight and the weights of brain, spleen, lungs, thymus and kidneys . Exposure to CCl(4) produced minimal effect on differential hematological parameters; however, it produced a significant increase in serum glutamic-pyruvic transaminase (SGPT) levels in all dose groups while other serum chemistries showed sporadic increases, primarily at the dose level of 1000 mg/kg . Exposure to CCl(4) produced a decreased humoral immune response; the IgM antibody forming cell (AFC) response to sheep red blood cells (sRBC) was suppressed with the maximal decrease (45%) observed at the dose level of 1000 mg/kg . The IgM serum titer to sRBC was also reduced with a maximal decrease (54%) observed at the dose level of 500 mg/kg . Although exposure to CCl(4) had no effects on the mixed leukocyte response (MLR), cytotoxic T lymphocyte activity and natural killer (NK) cell activity, a decrease in both the absolute number and the percentage of CD4(+)CD8(-) at the dose level of 500 mg/kg was observed . The functional activity of the mononuclear phagocyte system was compromised as reflected by a decrease in the vascular clearance of (51)Cr-sRBC and a decrease in the uptake of (51)Cr-sRBC by the liver . Finally, in the two host resistance models evaluated, exposure to CCl(4) decreased host resistance to both Streptococcus pneumoniae and Listeria monocytogenes with greater susceptibility to the latter . Overall, these studies demonstrate that CCl(4) was immunosuppressive in female B6C3F1 mice.

Enzyme Microb Technol, 2000 Dec, 27(10), 784 - 788
Data and knowledge based experimental design for fermentation process optimization; Berkholz R et al.; A novel method for the sequential experimental design in order to optimize fed-batch fermentations was applied to a hyaluronidase fermentation by Streptococcus agalactiae . A Lambda-optimal design was introduced to minimize the model parameter estimation error and to maximize the performance of the fermentation process . The method employs hybrid models that contain mechanistic, fuzzy and neural network components.

J Oral Maxillofac Surg, 2000 Dec, 58(12), 1347 - 52; discussion 1353
Cervical necrotizing fasciitis of odontogenic origin: a report of 11 cases; Tung-Yiu W et al.; PURPOSE: Although most cases of cervical necrotizing fasciitis (CNF) are odontogenic in origin, reports of this disease in the dental literature are sparse . The purpose of this study was to review the cases treated on our service, and to analyze the features of this disease and the responses to management, to supplement the understanding of this relatively rare and life-threatening disease . PATIENTS AND METHODS: All cases of infection admitted to the OMS service in a period of 10.5 years were studied retrospectively . The diagnosis of CNF was established by the findings on surgical exploration and histologic examination . The patients' age, sex, medical status, causes of the infection, bacteriology, computed tomography scan findings, surgical interventions, complications, survival, and other clinical parameters were reviewed . RESULTS: A total of 422 cases of infection were admitted, and 11 cases of cervical necrotizing fasciitis were found . The incidence of CNF was 2.6% among the infections hospitalized on the OMS service . There were 7 male and 4 female patients . Eight patients were older than 60 years of age . Seven patients had immunocompromising conditions, including diabetes mellitus in 4, concurrent administration of steroid in 2, uremia in 1, and a thymus carcinoma in 1 . All patients showed parapharyngeal space involvement; four also showed retropharyngeal space involvement . Gas was found in the computed tomography scan in 6 patients, extending to cranial base in 3 of them . Anaerobes were isolated in 73% of the infections, whereas Streptococcus species were uniformly present . All patients received 1 or more debridements . Major complications occurred in 4 patients, including mediastinitis in 4, septic shock in 2, lung empyema in 1, pleural effusion in 2, and pericardial effusion in 1 . All major complications developed in the immunocompromised patients, leading to 2 deaths . CONCLUSION: The mortality rate in this study was 18% . Early surgical debridement, intensive medical care, and a multidisciplinary approach are advocated in the management of CNF.

Eur J Clin Microbiol Infect Dis, 2000 Oct, 19(10), 755 - 8
Macrolide resistance phenotypes and genotypes in French clinical isolates of Streptococcus pneumoniae . Observatoire de Normandie du Pneumocoque; Angot P et al.; The aim of this study was to analyze the mechanisms of macrolide resistance in French clinical isolates of Streptococcus pneumoniae . A total of 838 strains of pneumococci were isolated in 1997 in Normandy, a region of western France, by 19 microbiology laboratories . Fifty-three percent had displayed diminished susceptibility to penicillin G and 50% were resistant to erythromycin . From this collection, 92 penicillin-intermediate or -resistant and 18 penicillin-susceptible strains resistant to erythromycin were studied . The presence of erm genes coding for ribosomal methylases and of mefE-like genes responsible for macrolide efflux was screened by a multiplex polymerase chain reaction and confirmed by DNA/DNA hybridization . Of the 110 strains studied, 108 were cross-resistant to erythromycin, spiramycin and clindamycin, including 105 strains containing ermB-related genes and three strains that contained a combination of ermB- and mefE-related genes . Two strains apparently susceptible to clindamycin but resistant to spiramycin also contained ermB-related genes . No strain was resistant to erythromycin alone or contained only a mef-like gene . Therefore, resistance to erythromycin is mostly related to ribosomal methylation in this region of France.

Indian J Ophthalmol, 2000 Jun, 48(2), 123 - 8
Spectrum of aetiological agents of postoperative endophthalmitis and antibiotic susceptibility of bacterial isolates; Anand AR et al.; PURPOSE: To determine the spectrum of infectious agents of postoperative endophthalmitis, the relationship with the time of onset of symptoms after surgery and the antibiotic susceptibilities of the aerobic bacterial isolates . METHODS: A retrospective review of microbiological records from January 1995 to December 1998 yielded 173 isolates from intraocular specimen of 170 patients with culture-proven postoperative endophthalmitis . Antibiotic susceptibility of these isolates was determined for various ocular antibiotics using the Kirby-Bauer disk-diffusion test . Based on the time of onset of illness, clinical presentation was classified into acute, delayed and chronic . RESULTS: Among 170 cases, 71 (41.7%) were attributable to gram-negative, 64 (37.6%) to gram-positive bacteria, and 37 (21.8%) to fungi . Gram-negative bacteria included P . aeruginosa (29;17.1%), other Pseudomonas spp (15;8.8%), non-fermenters (18;10.6%) and others (10;5.8%) . Among these, 40 of 72 (55.5%) were sensitive to gentamicin, 47 of 72 (65.2%) to cefotaxime, 47 of 69 (68.1%) to amikacin, 52 of 71 (73.2%) to ciprofloxacin, and 25 of 40 (62.5%) to ceftazidime . The gram-positive bacteria included S . epidermidis (22;12.9%), S . aureus (13;7.6%), P . acnes (10;5.9%), Enterococcus spp (4;2.3%), Streptococcus spp (7;4.1%) and others (8;4.8%) . Among these, 41 of 53 (77.3%) were sensitive to gentamicin, 47 of 53 (88.6%) to cefotaxime, 46 of 52 (88.4%) to ciprofloxacin, 38 of 41 (92.6%) to cefazolin and 27 of 37 (72.9%) to ceftazidime . All gram-positive bacteria were sensitive to vancomycin . CONCLUSION: In this large series of postoperative endophthalmitis, gram-negative bacilli followed by fungi accounted for the largest number of cases . A high degree of resistance of gram-negative bacilli to gentamicin, cefotaxime, amikacin and ceftazidime was recorded.

J Mol Evol, 2000 Dec, 51(6), 520 - 31
Structural evidence for the evolution of pyrogenic toxin superantigens; Mitchell DT et al.; Pathogenic bacteria have evolved a wide variety of toxins to invade and attack host organisms . In particular, strains of the bacteria Staphylococcus aureus and Streptococcus pyogenes produce a family of pyrogenic toxin superantigens (PTSAgs) that can cause illness, e.g., toxic shock syndrome, or synergize with a number of other immune system disorders . The PTSAgs are all similar in size and have a conserved two-domain tertiary fold despite minimal amino acid sequence identity . The tertiary structure of PTSAg domain 1 is similar to the immunoglobulin binding motif of streptococcal proteins G and L . PTSAg domain 2 resembles members of the oligosaccharide/oligonucleotide binding fold family that includes the B subunits of the AB(5) heat-labile enterotoxins, cholera toxin, pertussis toxin, and verotoxin . The strong structural homology between the pyrogenic toxins and other bacterial proteins suggests that the PTSAgs evolved through the recombination of two smaller beta-strand motifs.

Vaccine, 2000 Nov 22, 19(7-8), 850 - 61
Preparation and preclinical evaluation of experimental group B streptococcus type III polysaccharide-cholera toxin B subunit conjugate vaccine for intranasal immunization; Shen X et al.; Streptococcus group B (GBS) is usually carried asymptomatically in the vaginal tract of women and can be transferred to the newborn during parturition . Serum antibodies to the capsular polysaccharide (CPS) can prevent invasive diseases, whereas immunity acting at the mucosal surface may be more important to inhibit the mucosal colonization of GBS and thus the risk of infection for the newborn . We prepared different GBS type III CPS-protein conjugate vaccines and evaluated their systemic and mucosal immunogenicity in mice . GBS type III CPS was conjugated to tetanus toxoid (TT) or recombinant cholera toxin B subunit (rCTB) either directly or to rCTB indirectly via TT . The conjugation was performed by different methods: (1) CPS was coupled to TT with 1-ethyl-3 (3-dimethylaminopropyl)-carbodiimide (EDAC), using adipic acid dihydrazide (ADH) as a spacer; (2) CPS was conjugated with rCTB using reductive amination; or, (3) N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) was used to bind rCTB to the TT of the CPS-TT conjugate . Mice were immunized with these conjugates or purified CPS by subcutaneous (s.c.) and intranasal (i . n.) routes . Antibodies to GBS III in serum, lungs and vagina were measured with ELISA . All of the CPS-protein conjugates were superior to unconjugated CPS in eliciting CPS-specific immune responses in serum and mucosal tissue extracts . The conjugates, when administrated s.c., induced only IgG responses in serum, lung and vagina, while i.n . vaccination also elicited IgA responses in the lungs and vagina . The CPS-TT conjugate administrated i.n . induced a strong serum IgG, but only a weak mucosal IgA response, while the CPS-rCTB conjugate elicited high IgG as well as IgA antibodies in the lungs after i.n . immunization . GBS III CPS-TT conjugated with rCTB produced a strong systemic and local anti-CPSIII response after i.n . administration . Co-administration of CT as adjuvant enhanced the anti-CPS systemic and mucosal immune responses further after i.n . administration with the CPS conjugates . These findings indicate that: (i) i.n . immunization with GBS CPS-protein conjugates was more effective than s.c immunization for stimulating serum as well as mucosal immune responses; (ii) rCTB as a carrier protein for GBS III CPS could markedly improve the mucosal immune response; and (iii) the experimental GBS type III CPS conjugates containing rCTB should be investigated as mucosal vaccine to prevent GBS infection in humans.

Int J Pediatr Otorhinolaryngol, 2000 Dec 1, 56(2), 129 - 34
Prevention of acute mastoiditis: fact or fiction?
Linder TE, Briner HR, Bischoff T.
Acute mastoiditis is the most common complication of acute otitis media (AOM) . In recent years routine antibiotic treatment for acute middle ear infections was questioned and even abandoned in some countries . The goal of our study was to investigate the influence of antibiotic treatment on the occurrence and clinical outcome of acute mastoiditis and to analyze the bacteriological findings . A retrospective case record study of 48 patients with 50 episodes of acute mastoiditis hospitalized at our tertiary-care center between 1992 and 1999 was performed . Twenty-three patients (48%) received antibiotic treatment before admission whereas 25 (52%) did not . The group of patients without antibiotic pretreatment were younger (mean, 6 years) than patients with antibiotics (mean, 18 years) and their referral was delayed . The most common isolated single pathogen was Streptococcus pneumoniae . All pneumococci were sensitive to penicillin . Acute mastoiditis may be the first clinical sign of a middle ear infection, especially in very young children . Adequate antibiotic pretreatment cannot invariably prevent the development of acute mastoiditis even in the absence of penicillin resistant pathogens.

Mol Microbiol, 2000 Nov, 38(4), 867 - 78
Cross-regulation of competence pheromone production and export in the early control of transformation in Streptococcus pneumoniae; Martin B et al.; Two operons, comAB and comCDE, play a key role in the co-ordination of spontaneous competence development in cultures of Streptococcus pneumoniae . ComAB is required for export of the comC-encoded competence-stimulating peptide (CSP) . Upon CSP binding, the histidine kinase ComD activates ComE, its cognate response regulator, required for autoinduction of comCDE and for induction of the late competence genes . To understand better the early control of competence development, mutants upregulating comCDE (ComCDEUP) were isolated using a comC-lacZ transcriptional fusion . Mutants were generated by polymerase chain reaction mutagenesis of the comCDE region and by in vitro transposon mutagenesis of the chromosome . Both types of ComCDEUP mutants exhibited similar phenotypes . They differed from wild type in displaying trypsin-resistant transformation, competence under acid growth conditions and expression of comCDE under microaerobiosis; increased production of CSP in the mutants could account for the various phenotypes . The ComCDEUP transposon mutations included four independent insertions in the ciaR gene, which encodes the response regulator of a two-component system previously found to affect competence, and two immediately upstream of the comAB operon . The latter two resulted in comAB overexpression, indicating that CSP export is rate limiting . Among comDE point mutations, a single amino acid change in ComD (T233I) conferred constitutive, CSP-independent competence and resulted in comAB overexpression, providing support for the hypothesis that ComE regulates comAB; a ComE mutant (R120S) exhibited altered kinetics of competence shut-off . Collectively, these data indicate that pheromone autoinduction, cross-regulation of the comAB and comCDE operons and, possibly, competence shut-off contribute to the early control of competence development in S . pneumoniae . They argue for a metabolic control of competence, mediated directly or indirectly by CiaR, and they suggest that both comAB and comCDE are potential targets for regulation.

Clin Infect Dis, 2001 Jan, 32(1), 9 - 16 Epub 2000 Dec 08.
Spinal epidural abscesses in children: a 15-year experience and review of the literature; Auletta JJ et al.; We reviewed medical records and laboratory and diagnostic evaluations for 8 pediatric patients with spinal epidural abscesses who were treated during the last 15 years at our institution . Staphylococcus aureus was isolated from 5 of 8 epidural abscesses, including 2 abscesses with methicillin-resistant S . aureus . Unusual isolates were group B Streptococcus in a patient with chronic vesicouretral reflux associated with the posterior urethral valves and Aspergillus flavus in a patient with acute myelogenous leukemia . An analysis incorporating our results and a review of the English-language literature about abscesses in children and adults revealed differences related to age . Abscesses in children were more posterior in epidural location, had greater spinal column extension, and were associated with more favorable clinical outcomes than were abscesses in adults . Magnetic resonance imaging is the diagnostic procedure of choice; however, radionuclide bone scans should be considered for associated distant osteomyelitis in children . Prompt diagnosis and combined medical and surgical treatment remain the cornerstones for the prevention of adverse outcomes.

Org Lett, 2000 Dec 14, 2(25), 4013 - 5
Asymmetric synthesis of quaternary centers . Total synthesis of (-)-malyngolide; Trost BM et al.; {structure} The deracemization of 3-nonyl-3,4-epoxybut-1-ene with Pd(0) in the presence of chiral ligands using p-methoxybenzyl alcohol as a nucleophile proceeds regio- and enantioselectively to form the monoprotected vinylglycidol in 99% ee . This chiral building block was converted in seven steps to (-)-malyngolide, an antibiotic showing significant activity against Mycobacterium smegmatis and Streptococcus pyogenes . An interesting aspect involves controlling the diastereoselectivity of protonation of an enolate via a distal hydroxyl group.

J Pediatr Health Care, 2000 Nov-Dec, 14(6), 264 - 9
Early-onset neonatal group B streptococcal infection: implications for practice; Parks DK et al.; Group B streptococcus (GBS) is the leading bacterial infection associated with morbidity and mortality of newborns in the United States . Most neonatal infections can be prevented through the use of intrapartum antimicrobial prophylaxis in women who are at increased risk for transmitting infection to their newborns . However, prevention strategies have not been implemented widely or consistently, and the incidence of neonatal GBS disease has not declined . An understanding of GBS epidemiology, clinical presentation, and prevention strategies enhances the PNP's decision-making skills in the nursery and strengthens the PNP's ability to evaluate and compare new approaches to GBS prevention.

J Invertebr Pathol, 2000 Nov, 76(4), 233 - 41
Apolipophorin-III and the interactions of lipoteichoic acids with the immediate immune responses of Galleria mellonella; Halwani AE et al.; We investigated the effects of lipoteichoic acids, surface components of Gram-positive bacteria, on the hemocytes and phenoloxidase activity in last instar Galleria mellonella larvae, as well as the binding of apolipophorin-III, an insect lipid-binding protein, to lipoteichoic acids . Binding of apolipophorin-III to lipoteichoic acid was studied using an assay based on 1,9-dimethylmethylene blue . Apolipophorin-III bound the lipoteichoic acids from Bacillus subtilis, Enterococcus hirae, and Streptococcus pyogenes and to intact cells of E . hirae . E . hirae lipoteichoic acid promoted the binding of apolipophorin-III to the cells of this species . All lipoteichoic acids tested caused a dose- and time-dependent drop in the total counts of hemocytes and, depending on the species of lipoteichoic acid, partial or complete depletion of plasmatocytes . Granulocyte counts were not affected . Apolipophorin-III prevented partially the loss of plasmatocytes due to B . subtilis lipoteichoic acid . All three lipoteichoic acids studied activated phenoloxidase in vitro; injections of B . subtilis lipoteichoic acid into the larvae elevated the phenoloxidase activity, whereas injections of E . hirae or S . pyogenes lipoteichoic acid, or apolipophorin-III alone, suppressed it . Apolipophorin-III decreased the activation of phenoloxidase by B . subtilis lipoteichoic acid.

J Infect Dis, 2001 Jan 15, 183(2), 253 - 260 Epub 2000 Dec 08.
Serum samples from infants vaccinated with a pneumococcal conjugate vaccine, PncT, protect mice against invasive infection caused by Streptococcus pneumoniae serotypes 6A and 6B; Saeland E et al.; Streptococcus pneumoniae serogroup 6 is an important cause of respiratory tract disease worldwide . Vaccination with 6B polysaccharide induces antibody response to the cross-reacting serotype 6A, but the protective capacity of 6A antibodies induced in infants remains unknown . In this study, passive immunization with serum samples obtained from infants vaccinated with an octavalent polysaccharide protein conjugate vaccine, PncT, protected mice against bacteremia and/or lung infection caused by intranasal challenge with serotypes 6B and 6A . Protective infant serum samples had significantly higher serotype-specific IgG levels and opsonic activity than did nonprotective serum samples . The protective level to either serotype was approximately 1 microg of specific IgG antibodies injected per mouse (corresponding to approximately 0.3 microg/mL) . The protection was strongly related to opsonophagocytic antibody levels measured in vitro . These results demonstrate that PncT induces antibodies in infants that protect mice against invasive disease caused by the homologous serotype and by the cross-reacting serotype 6A.

Enferm Infecc Microbiol Clin, 2000 Aug-Sep, 18(7), 314 - 8
{Resistance to penicillin and other antimicrobials in 301 clinical isolates of Streptococcus pneumoniae}; Navarro C et al.; BACKGROUND: The aim of this study was to assess the susceptibility to penicillin of Streptococcus pneumoniae clinical strains and to analyze the association between penicillin resistance and cefotaxime and cefixime activity in S . pneumoniae isolates with decreased sensitivity to penicillin . METHODS: 301 S . pneumoniae clinical strains were isolated from patients during 1995-1996 . Susceptibility to penicillin, cefotaxime, cefepime, erythromycin, chloramphenicol, tetracycline, cotrimoxazole and ciprofloxacin were studied . RESULTS: 38.2% isolates were penicillin-susceptible and 61.8% were penicillin-resistant; 20.6% showed high-level resistance . Resistance rates to erythromycin, chloramphenicol, tetracycline, cotrimoxazole and ciprofloxacin were, respectively, 30.9, 30.2, 40.9, 66.4, and 13.3% overall, and 54.8, 54.8, 61.3, and 93.5% in the 62 strains with high-level resistance to penicillin . Strains resistant to cefotaxime and cefepime were 13.9 and 14.9%, respectively . MIC50 and MIC90 for cefotaxime and cefepime in penicillin-resistant strains were 0.5 and 1 mg/ml . CONCLUSIONS: A high proportion of S . pneumoniae isolates showed resistance to penicillin, in agreement with other Spanish reports . Moreover, resistance to penicillin was significantly associated (p < 0.001) with resistance to erythromycin, chloramphenicol, tetracycline and cotrimoxazole, but not with ciprofloxacin . MIC50 and MIC90 for cefotaxime and cefepime were similar, and lower than those for penicillin in penicillin-resistant pneumococci strains.

Rozhl Chir, 2000 Sep, 79(9), 414 - 7
{Streptococcus pneumoniae--an infectious agent in coxitis}; Pospisil M et al.; Based on their own observation the authors describe pneumococcal coxitis in a young man . With regard to the atypical clinical picture of the disease the authors draw attention to the necessity to consider in the differential diagnosis of pain in the inguinal and coxal area also infectious arthritis . This concerns in the first place physicians in the first line of contact.

Diagn Microbiol Infect Dis, 2000 Nov, 38(3), 177 - 9
In vitro activity of GAR-936 against vancomycin-resistant enterococci, methicillin-resistant Staphylococcus aureus and penicillin-resistant Streptococcus pneumoniae; Patel R et al.; We report the activity of the new glycylcycline antimicrobial agent GAR-936 against 37 clinical isolates of vancomycin-resistant enterococci (including organisms carrying the vanA, vanB, vanC-1, and vanC-2/3 genes), 26 clinical isolates of methicillin-resistant S . aureus and 30 clinical isolates of high-level penicillin-resistant S . pneumoniae . All isolates of vancomycin-resistant enterococci, methicillin-resistant S . aureus, and penicillin-resistant S . pneumoniae were inhibited by < or = 1, < or = 2, or < or = 0.25 microg/ml of GAR-936, respectively . Time kill experiments using vancomycin-resistant enterococci did not demonstrate synergy or antagonism between 2 microg/ml of GAR-936 and 0.25 microg/ml of quinupristin/dalfopristin.

Diagn Microbiol Infect Dis, 2000 Nov, 38(3), 171 - 2
Group A streptococcal appendicitis in a patient with AIDS; Tufariello JM et al.; A man with AIDS developed appendicitis and bacteremia caused by Group A streptococcus, neither of which is considered an opportunistic infection . Group A streptococcus is rarely implicated in appendicitis in children and has not previously been reported in an adult . Immunodeficiency might have predisposed the patient to this unusual infection.

Diagn Microbiol Infect Dis, 2000 Nov, 38(3), 151 - 7
The use of Monte Carlo simulation to examine pharmacodynamic variance of drugs: fluoroquinolone pharmacodynamics against Streptococcus pneumoniae; Ambrose PG et al.; BACKGROUND: For fluoroquinolones, AUC:MIC ratios correlate with maximal bacterial eradication in in vitro models of infection and favorable cure rates in humans with respiratory tract infection . Inter-subject pharmacokinetic and MIC variability may impact the probability of attaining optimal AUC:MIC ratios and hence favorable clinical outcome . METHODS: Monte Carlo simulation was utilized to estimate the probability of attaining AUC:MIC ratios of 30, 40, 50, 60, 70, 80, 90, 100, 110 and 120 using AUC values from patients treated with either gatifloxacin or levofloxacin and microbiologic activity against S . pneumoniae observed in 1997 SENTRY Antimicrobial Surveillance Program . RESULTS: The probability curves for 5000 patient simulations were plotted . The median AUC:MIC ratios were 120 for gatifloxacin and 50.5 for levofloxacin . The probability of attaining AUC:MIC ratios of 30, 50, 70 and 100 for gatifloxacin were 94%, 86%, 78% and 62%, and for levofloxacin were 80%, 51%, 31% and 17%, respectively . CONCLUSION: Gatifloxacin has a higher probability of achieving target AUC:MIC ratios than levofloxacin . Monte Carlo simulation, using patient-based AUC and MIC distributions, may have implications for selection of optimal antibiotics for the empiric treatment of infections . Moreover, Monte Carlo simulation may have utility in the determination of MIC breakpoints.

Neurosci Lett, 2000 Dec 22, 296(2-3), 137 - 40
Spatial memory and learning deficits after experimental pneumococcal meningitis in mice; Wellmer A et al.; Survivors of bacterial meningitis frequently suffer from long-term sequelae, particularly from learning and memory deficits . For this reason, spatial memory and learning was studied in a mouse model of ceftriaxone-treated Streptococcus pneumoniae meningitis . Persistent deficits of spatial learning despite normal motor function were observed in mice infected with 10(4) colony-forming units (CFU) in 25 microl of saline into the right forebrain in comparison to mice treated with an equal amount of saline . Survivors of meningitis performed significantly worse in memorizing a hidden platform in a Morris water maze . After 2 weeks, the difference between post-meningitis and control mice diminished . Yet, when the platform was moved after 180 days, learning of the new location was still strongly impaired in mice surviving meningitis.

Int Microbiol, 1999 Sep, 2(3), 169 - 76
Functional organization of the gene cluster involved in the synthesis of the pneumococcal capsule; Garcia E et al.; Streptococcus pneumoniae is a major human pathogen and its capsular polysaccharide has been shown to be the main virulence factor . The molecular organization of the genes governing the formation of this capsule was not studied until the 1990s . The capsular clusters (cap) of eight of the 90 known pneumococcal types have now been studied . The cap operon, located between the dexB and aliA genes, is arranged as a central region comprising the genes coding for the specific-type polysaccharide, flanked by open reading frames that are mostly common to all of the serotypes . The biochemical functions of 24 genes required for capsular polysaccharide biosynthesis have been elucidated but the precise role of the flanking regions in capsular formation is unknown . The natural genetic transformation characteristic of pneumococci, the arrangement of the cap locus and the abundance of transposable elements at this locus favor the genetic variability of the capsule in this microorganism . These well-documented observations together with the finding that some genes located outside the cap cluster may also participate in capsule formation increase the complexity of pneumococcal infection control.

Int Microbiol, 1999 Mar, 2(1), 23 - 8
Construction of a new Streptococcus pneumoniae-Escherichia coli shuttle vector based on the replicon of an indigenous pneumococcal cryptic plasmid; Munoz R et al.; The nucleotide sequence of a cryptic plasmid (pRMG1) isolated from a type 1 Streptococcus pneumoniae has been determined and two recombinant plasmids, pRMGE1 and pRMGE2, bearing the pRMG1 replicon have been constructed . pRMGE2 is a shuttle vector for Escherichia coli and S . pneumoniae . The important characteristics of this cloning vector are: a size of 5.5 kb including a 1.4 kb fragment of pRMG1 (containing a double-stranded replication origin and an open reading frame encoding a putative replication initiation protein), a multicloning site, two antibiotic resistance markers for selection of plasmid containing cells, and blue-white colony screening in E . coli for identification of insert-containing plasmids.

Syst Appl Microbiol, 2000 Oct, 23(3), 330 - 2
Identification of methionine-processed HPr in the equine pathogen Streptococcus equi; Sutcliffe IC et al.; Using preparative electrophoresis, a low molecular weight protein has been partially purified from a cell extract of the equine pathogen Streptococcus equi susp . equi . N-terminal sequence analysis and Western blotting revealed the protein to be HPr, a central component of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) . Interestingly, the only form of the HPr protein detected in S . equi was one with the amino-terminal methionine removed, a modification that has previously been associated with surface localization of streptococcal HPr proteins.

Syst Appl Microbiol, 2000 Oct, 23(3), 325 - 9
Characterization of acid phosphatase activities in the equine pathogen Streptococcus equi; Hamilton A et al.; Acid phosphatases hydrolyse phosphomonoesters at acidic pH in a variety of physiological contexts . The recently defined class C family of acid phosphatases includes the 32 kDa LppC lipoprotein of Streptococcus equisimilis . To define further the distribution of acid phosphatases in the genus Streptococcus we have examined the equine pathogens Streptococcus equi subsp . equi and Streptococcus equi subsp . zooepidemicus . Whole cell assays indicated that these organisms possess two acid phosphatases with activity optima at pH 5.0 and pH 6.0-6.5 and that only the former of these was, like LppC, resistant to EDTA . Western blotting with a polyclonal anti-LppC antiserum revealed the presence of a cross-reactive 32 kDa protein in both organisms . The cross-reactive protein in S . equi was shown to be a surface accessible lipoprotein as its processing was inhibited by the antibiotic globomycin and it was released from whole cells by treatment with trypsin . The presence of DNA sequences homologous to the S . equisimilis lppC gene were confirmed by PCR . These data strongly suggest that Streptococcus equi subsp . equi and Streptococcus equi subsp . zooepidemicus produce a lipoprotein acid phosphatase homologous to LppC of S . equisimilis.

Biochemistry, 2000 Dec 12, 39(49), 14993 - 5001
The structure of UDP-N-acetylglucosamine 2-epimerase reveals homology to phosphoglycosyl transferases; Campbell RE et al.; Bacterial UDP-N-acetylglucosamine 2-epimerase catalyzes the reversible epimerization at C-2 of UDP-N-acetylglucosamine (UDP-GlcNAc) and thereby provides bacteria with UDP-N-acetylmannosamine (UDP-ManNAc), the activated donor of ManNAc residues . ManNAc is critical for several processes in bacteria, including formation of the antiphagocytic capsular polysaccharide of pathogens such as Streptococcus pneumoniae types 19F and 19A . We have determined the X-ray structure (2.5 A) of UDP-GlcNAc 2-epimerase with bound UDP and identified a previously unsuspected structural homology with the enzymes glycogen phosphorylase and T4 phage beta-glucosyltransferase . The relationship to these phosphoglycosyl transferases is very intriguing in terms of possible similarities in the catalytic mechanisms . Specifically, this observation is consistent with the proposal that the UDP-GlcNAc 2-epimerase-catalyzed elimination and re-addition of UDP to the glycal intermediate may proceed through a transition state with significant oxocarbenium ion-like character . The homodimeric epimerase is composed of two similar alpha/beta/alpha sandwich domains with the active site located in the deep cleft at the domain interface . Comparison of the multiple copies in the asymmetric unit has revealed that the epimerase can undergo a 10 degrees interdomain rotation that is implicated in the regulatory mechanism . A structure-based sequence alignment has identified several basic residues in the active site that may be involved in the proton transfer at C-2 or stabilization of the proposed oxocarbenium ion-like transition state . This insight into the structure of the bacterial epimerase is applicable to the homologous N-terminal domain of the bifunctional mammalian UDP-GlcNAc "hydrolyzing" 2-epimerase/ManNAc kinase that catalyzes the rate-determining step in the sialic acid biosynthetic pathway.

Eur J Biochem, 2000 Dec, 267(24), 7147 - 57
Structures of two cell wall-associated polysaccharides of a Streptococcus mitis biovar 1 strain . A unique teichoic acid-like polysaccharide and the group O antigen which is a C-polysaccharide in common with pneumococci; Bergstrom N et al.; The cell wall of Streptococcus mitis biovar 1 strain SK137 contains the C-polysaccharide known as the common antigen of a closely related species Streptococcus pneumoniae, and a teichoic acid-like polysaccharide with a unique structure . The two polysaccharides are different entities and could be partially separated by gel chromatography . The structures of the two polysaccharides were determined by chemical methods and by NMR spectroscopy . The teichoic acid-like polymer has a heptasaccharide phosphate repeating unit with the following structure: The structure neither contains ribitol nor glycerol phosphate as classical teichoic acids do, thus we have used the expression teichoic acid-like for this polysaccharide . The following structure of the C-polysaccharide repeating unit was established: where AAT is 2-acetamido-4-amino-2,4, 6-trideoxy-D-galactose . It has a carbohydrate backbone identical to that of one of the two structures of C-polysaccharide previously identified in S . pneumoniae . C-polysaccharide of S . mitis is characterized by the presence, in each repeating unit, of two residues of phosphocholine and both galactosamine residues in the N-acetylated form . Immunochemical analysis showed that C-polysaccharide constitutes the Lancefield group O antigen . Studies using mAbs directed against the backbone and against the phosphocholine moiety of the C-polysaccharide revealed several different patterns of these epitopes among 95 S . mitis and Streptococcus oralis strains tested and the exclusive presence of the group O antigen in the majority of S . mitis biovar 1 strains.

Vet Rec, 2000 Nov 11, 147(20), 563 - 7
Investigations towards an efficacious and safe strangles vaccine: submucosal vaccination with a live attenuated Streptococcus equi; Jacobs AA et al.; As part of a search for a safe and efficacious strangles vaccine, several different vaccines and different vaccination routes were tested in foals . The degree of protection was evaluated after an intranasal challenge with virulent Streptococcus equi by clinical, postmortem and bacteriological examinations . Inactivated vaccines containing either native purified M-protein (500 microg per dose) or whole S equi cells (10(10) cells per dose) administered at least twice intramuscularly at intervals of four weeks, did not protect against challenge . Different live attenuated S equi mutants administered at least twice at intervals of four weeks by the intranasal route were either safe but not protective or caused strangles . In contrast, a live attenuated deletion mutant administered intramuscularly, induced complete protection but also induced unacceptable local reactions at the site of vaccination . Submucosal vaccination in the inner side of the upper lip with the live attenuated mutant at > or =10(8) colony-forming units per dose, appeared to be safe and efficacious in foals as young as four months of age . The submucosal vaccinations caused small transient swellings that resolved completely within two weeks, and postmortem no vaccine remnants or other abnormalities were found at the site of vaccination.

Handchir Mikrochir Plast Chir, 2000 Sep, 32(5), 343 - 6
{Chronic paronychia and synovialitis of extensor tendons due to Mycobacterium marinum . Is diagnosis or treatment the problem?}; Witthaut J et al.; Most infections of the upper extremity are caused by staphylococcus or streptococcus and respond well to beta-lactam antibiotics . Hand surgeons should be aware of the possible diagnosis of Mycobacterium marinum infection: 90% of the lesions are found in the upper extremity . We present a case of a chronic, cutaneous lesion of the right middle finger with synovialitis of the extensor tendons observed in a 35-year-old woman . Routine cultures from tissue of the infected finger led to the diagnosis of paronychia due to staphylococcus aureus . Despite surgical and antibacterial treatment, the lesion persisted and the patient developed multiple raised, non-tender satellite lesions to the right hand and elbow . Based on the clinical aspect and a detailed history (she kept fish and had suffered a chicken bone stab to her middle finger 12 weeks earlier), we suspected a Mycobacterium marinum infection . Tissue was obtained mainly by synovialectomy . Culture of the biopsy tissue for Mycobacterium marinum confirmed the diagnosis . The patient responded to a triple therapy (rifabutin, ethambutol and clarithromycin) and had an uncomplicated recovery . The importance of a high index of suspicion, adequate examination and a complete patient's history for a correct diagnosis is stressed . Culture for Mycobacterium marinum is not routinely performed and ought to be initiated once an infection is suspected . We also discuss the best timing for the onset of medical treatment.

J Neurol Sci, 2000 Dec 15, 182(1), 36 - 44
Adult bacterial meningitis in Southern Taiwan: epidemiologic trend and prognostic factors; Lu CH et al.; In two investigative phases over a 13.5-year study period (January 1986-June 1999), 202 adult patients with culture-proven bacterial meningitis were enrolled in this study . In order to determine the epidemiologic trend, prognostic factors and therapeutic results for this disease . Klebsiella pneumoniae (K . pneumoniae), Pseudomonas aeruginosa, and Streptococcus pneumoniae were the three most commonly revealed pathogens, accounting for about 48% of the episodes . Although there was a change in relative frequency for the pathogens, K . pneumoniae remained the most prevalent during the two periods studied (January 1986-December 1992 and January 1993-June 1999) . Multiantibiotic resistant strains have been in evidence since their appearance in 1994, with most of our patients acquiring their infection nosocomially . The overall mortality rates during the two periods were 40% and 34%, respectively . In stepwise logistic regression analysis, only initial conscious level, appropriate antibiotic therapy and septic shock were independently associated with mortality, after adjustment for other potentially confounding factors . Initial empirical antibiotics with both third-generation cephalosporin and penicillin G, should be considered for the majority of meningitis cases resulting from infection with Gram-negative bacilli and streptococcal species . Besides the evolution of newer pathogens, there has been increasing incidence for nosocomially acquired bacterial meningitis for patients postneurosurgery, with the emergence of resistant strains presenting a therapeutic challenge in recent years . Vancomycin and imipenem/cilastatin should be considered as the initial empirical antibiotics of choice for the treatment of this special group of patients.

J Antimicrob Chemother, 2000 Dec, 46(6), 973 - 80
Antagonism between penicillin and erythromycin against Streptococcus pneumoniae in vitro and in vivo; Johansen HK et al.; The combination of beta-lactam antibiotics and macrolides is often recommended for the initial empirical treatment of acute pneumonia in order to obtain activity against the most important pathogens . Theoretically, this combination may be inexpedient, as the bacteriostatic agent may antagonize the effect of the bactericidal agent . In this study, the possible interaction between penicillin and erythromycin was investigated in vitro and in vivo against four clinical isolates of Streptococcus pneumoniae with MICs of penicillin ranging from 0.016 to 0.5 mg/L and of erythromycin from 0 . 25 to >128 mg/L . In vitro time-kill curves were generated with clinically relevant concentrations of penicillin (10 mg/L) and erythromycin (1 mg/L), either individually or in combination . Antagonism between penicillin and erythromycin was observed for the four isolates . In vivo interaction was investigated in the mouse peritonitis model . After intraperitoneal inoculation, penicillin and erythromycin were given either individually or in combination . For two of the four isolates, mortality was significantly higher in the groups treated with the combination of penicillin and erythromycin than in the groups treated with penicillin alone {32/36 (86%) vs . 3/12 (25%), P<0.05; and 24/36 (67%) vs . 3/12 (25%), P<0.05, respectively} . Using the mouse peritonitis model, in vivo time-kill curves showed that there was antagonism between erythromycin and penicillin for the examined isolate . The antagonism demonstrated in vitro and in vivo between penicillin and erythromycin suggests that ss-lactam antibiotics and macrolides should not be administered together unless pneumococcal infection is ruled out.

J Antimicrob Chemother, 2000 Dec, 46(6), 959 - 64
Streptococcus pyogenes resistance to erythromycin in relation to macrolide consumption in Spain (1986-1997); Granizo JJ et al.; The relationship between Streptococcus pyogenes resistance to erythromycin and macrolide consumption in Spain was studied . Erythromycin resistance was highly correlated with the consumption of total macrolides (r = 0.88, P<0.01) . When macrolides were grouped into posological subgroups according to their pharmacokinetic and pharmacodynamic properties and analysed separately, erythromycin resistance appeared to be related mainly to those macrolides taken twice daily (bd) (r = 0.86, P<0.01) and those taken once daily (od) (r = 0.87, P<0.01), but not to those taken four (qds) or three times a day (tds) (r = -0.04, P: = 0.90) . A progressive increase in the erythromycin resistance curve was seen after the consecutive introduction of both bd and od macrolides, which contributed to the increase in the total macrolide consumption, replacing tds macrolide prescription . Although this ecological analysis cannot establish an unequivocal causal relationship between antibiotic consumption and S . pyogenes resistance, the data are consistent with the hypothesis that widespread use of macrolides, mainly of bd and od macrolides, resulted in an increased prevalence of S . pyogenes resistant to erythromycin in Spain.

J Antimicrob Chemother, 2000 Dec, 46(6), 909 - 15
In vitro development of resistance to ceftriaxone, cefprozil and azithromycin in Streptococcus pneumoniae; Nagai K et al.; Approval of ceftriaxone for the treatment of otitis media has led to fear of selection of resistant mutants owing to widespread use . To test this, we examined the ability of sequential subcultures in sub-MICs of ceftriaxone, cefprozil and azithromycin to select resistant mutants in 12 pneumococci . Daily subculturing was performed 50 times or until mutants with raised ceftriaxone, cefprozil or azithromycin MICs were selected . Of eight ceftriaxone-susceptible parents, ceftriaxone did not select for any resistant mutants, while cefprozil selected for four mutants (MICs 2-4 mg/L after 21-50 subcultures) . Among four ceftriaxone-resistant parents, subculturing in ceftriaxone selected for one stable mutant with raised ceftriaxone MIC (>16 mg/L after 21 subcultures) and subculturing in cefprozil selected for one mutant with raised cefprozil MIC (64 mg/L after 44 subcultures) . Mutations were observed in pbp2x and pbp1a . Among six azithromycin-susceptible parents, subculturing in azithromycin selected for five resistant mutants (MIC 0.5-32 mg/L after 10-42 passages) and among six azithromycin-resistant strains, subculturing selected for mutants with raised azithromycin MICs in all six strains (MIC 16-32 mg/L after 4-18 passages) . All azithromycin-resistant mutants derived from azithromycinsusceptible parents had mutations in domain V of 23S rRNA while all azithromycin-resistant parents and derived mutants had mefE . Single-step mutation rates among the 12 strains at the MIC ranged from 1.5 x 10(-6) to <6.2 x 10(-10) for ceftriaxone, >1.3 x 10(-5) to 8.9 x 10(-8) for cefprozil and >1.1 x 10(-6) to 6.7 x 10(-10) for azithromycin . Multi-step and single-step testing showed that ceftriaxone selected for resistant mutants less often than cefprozil and azithromycin.

J Antimicrob Chemother, 2000 Dec, 46(6), 905 - 8
In vitro activity of ketolides telithromycin and HMR 3004 against italian isolates of Streptococcus pyogenes and Streptococcus pneumoniae with different erythromycin susceptibility; Giovanetti E et al.; Two ketolides, telithromycin and HMR 3004, were evaluated for their in vitro activity against erythromycin-susceptible and -resistant strains of Streptococcus pyogenes and Streptococcus pneumoniae . On the basis of their resistance to macrolide, lincosamide and streptogramin (MLS) antibiotics, erythromycin-resistant test strains were assigned to the constitutive resistance (cMLS) phenotype, the inducible resistance (iMLS) phenotype or the M phenotype . iMLS S . pyogenes strains were further subdivided into the three recently described subtypes iMLS-A, -B and -C . Telithromycin and HMR 3004 were uniformly and highly active against pneumococci (regardless of their susceptibility or resistance to erythromycin and/or penicillin), erythromycin-susceptible S . pyogenes and erythromycin-resistant S . pyogenes strains of the M phenotype (in which resistance is mediated by an efflux system) or iMLS-B or -C phenotype (in which resistance is mediated by a methylase encoded by the ermTR gene) . Both ketolides were less active against erythromycin-resistant S . pyogenes strains with the cMLS phenotype or the iMLS-A subtype (where resistance is mediated by a methylase encoded by the ermAM gene), these strains ranging in phenotype from the upper limits of susceptibility to low-level resistant.

J Clin Microbiol, 2000 Dec, 38(12), 4548 - 53
Genetic relatedness within serotypes of penicillin-susceptible Streptococcus pneumoniae isolates; Overweg K et al.; The molecular epidemiological characteristics of all Streptococcus pneumoniae strains isolated in a nationwide manner from patients with meningitis in The Netherlands in 1994 were investigated . Restriction fragment end labeling analysis demonstrated 52% genetic clustering among these penicillin-susceptible strains, a value substantially lower than the percentage of clustering among Dutch penicillin-nonsusceptible strains . Different serotypes were found within 8 of the 28 genetic clusters, suggesting that horizontal transfer of capsular genes is common among penicillin-susceptible strains . The degree of genetic clustering was much higher among serotype 3, 7F, 9V, and 14 isolates than among isolates of other serotypes, i.e., 6A, 6B, 18C, 19F, and 23F . We further studied the molecular epidemiological characteristics of pneumococci of serotype 3, which is considered the most virulent serotype and which is commonly associated with invasive disease in adults . Fifty epidemiologically unrelated penicillin-susceptible serotype 3 invasive isolates originating from the United States (n = 27), Thailand (n = 9), The Netherlands (n = 8), and Denmark (n = 6) were analyzed . The vast majority of the serotype 3 isolates (74%) belonged to two genetically distinct clades that were observed in the United States, Denmark, and The Netherlands . These data indicate that two serotype 3 clones have been independently disseminated in an international manner . Seven serotype 3 isolates were less than 85% genetically related to the other serotype 3 isolates . Our observations suggest that the latter isolates originated from horizontal transfer of the capsular type 3 gene locus to other pneumococcal genotypes . In conclusion, epidemiologically unrelated serotype 3 isolates were genetically more related than those of other serotypes . This observation suggests that serotype 3 has evolved only recently or has remained unchanged over long periods.

J Clin Microbiol, 2000 Dec, 38(12), 4420 - 4
Bacterial colonization of disposable soft contact lenses is greater during corneal infiltrative events than during asymptomatic extended lens wear; Sankaridurg PR et al.; Microorganisms, especially gram-negative bacteria, are considered to play a role in the etiology of certain corneal infiltrative events (CIEs) observed during soft contact lens wear . This study explored the possibility of microbial colonization of soft contact lenses as a risk factor leading to CIEs . In a clinical trial conducted from March 1993 to January 1996, 330 subjects wore disposable soft contact lenses on a 6-night extended-wear and disposal schedule . During this period, 4,321 lenses (118 during CIEs; 4,203 during asymptomatic lens wear) were recovered aseptically and analyzed for microbial colonization . A greater percentage of lenses were free from microbial colonization during asymptomatic wear than during CIEs (42 versus 23%; P < 0.0001) . The incidence of gram-positive bacteria, gram-negative bacteria and fungi was greater during CIEs than during asymptomatic lens wear (P < 0.05) . During asymptomatic lens wear, gram-positive bacteria were isolated most frequently and were usually normal external ocular microbiota . Of the gram-positive bacteria, the incidence of Streptococcus pneumoniae was greater during CIE than during asymptomatic wear (7.6 versus 0.6%; P < 0 . 0001) . While gram-negative bacteria were seen in few cases during asymptomatic wear, their incidence during CIE in comparison to asymptomatic wear was substantial and significant (23.7 versus 3.8%; P < 0.0001) . Also, the level of colonization was high . Of CIEs, events of microbial keratitis, contact lens acute red eye, and asymptomatic infiltrative keratitis were associated with lens colonization with gram-negative bacteria or S . pneumoniae . Colonization of soft contact lenses with pathogenic bacteria, especially gram-negative bacteria and S . pneumoniae, appears to be a significant risk factor leading to CIE.

J Clin Microbiol, 2000 Dec, 38(12), 4367 - 72
Clonal groups of penicillin-nonsusceptible Streptococcus pneumoniae in Baltimore, Maryland: a population-based, molecular epidemiologic study; McEllistrem MC et al.; Few data are available on the molecular subtypes of all penicillin-nonsusceptible Streptococcus pneumoniae (PNSP) from a defined population base . Pulsed-field gel electrophoresis (PFGE), serotyping, and antibiotic susceptibility testing were performed for all available invasive PNSP isolates for which the penicillin (MIC) was > or =0.1 microg/ml from Baltimore, Md., during 1995-1996 (n = 143) . The dendrogram analysis of PFGE patterns included 32 distinct clonal groups . Six major clonal groups included two-thirds of the PNSP strains . Major clonal groups 2, 3, 4, and 6 strains were genetically related to four previously described international clones and were all multidrug resistant . Major clonal group 3 was genetically related to the Tennessee(23F)-4 clone and contained all four strains for which the penicillin MIC was 8 microg/ml . Most of the clonal group 1 and 5 strains had intermediate susceptibility to penicillin and were rarely multidrug resistant . The latter clonal groups represent two previously undescribed penicillin-intermediate pneumococcal clones . Clonal group homogeneity was greater for serotype 9V, 19A, and 23F strains than for serotype 6A, 6B, 14, and 19F strains . The classification of PNSP strains into clonal groups is essential for future population-based epidemiologic studies of PNSP.

J Clin Microbiol, 2000 Dec, 38(12), 4361 - 6
Molecular epidemiology of penicillin-nonsusceptible Streptococcus pneumoniae among children in Greece; Bogaert D et al.; A total of 145 penicillin-nonsusceptible Streptococcus pneumoniae strains were isolated from young carriers in Greece and analyzed by antibiotic susceptibility testing, serotyping, restriction fragment end labeling (RFEL), and penicillin-binding protein (PBP) genotyping . The serotypes 23A and 23F (54%), 19A and 19F (25%), 9V (5%), 15A, 15B, and 15C (4%), 6A and 6B (4%), and 21 (4%) were most prevalent in this collection . Fifty-three distinct RFEL types were identified . Sixteen different RFEL clusters, harboring 2 to 32 strains each, accounted for 82% of all strains . Eight of these genetic clusters representing 60% of the strains were previously identified in other countries . A predominant lineage of 66 strains (46%) harboring five RFEL types and the serotypes 19F and 23F was closely related to the pandemic clone Spain(23F)-1 (genetic relatedness of > or =85%) . Another lineage, representing 11 strains, showed close genetic relatedness to the pandemic clone France(9V)-3 . Another lineage of 8 serotype 21 strains was Greece specific since the RFEL types were not observed in an international collection of 193 genotypes from 16 different countries . Characterization of the PBP genes pbp1a, pbp2b, and pbp2x revealed 20 distinct PBP genotypes of which PBP type 1-1-1, initially observed in the pandemic clones 23F and 9V, was predominantly present in 11 RFEL types in this Greek collection of penicillin-nonsusceptible strains (55%) . Sixteen PBP types covering 52 strains (36%) were Greece specific . This study underlines the strong contribution of penicillin-resistant international clones to the prevalence and spread of penicillin-nonsusceptible pneumococci among young children in Greece.

J Craniofac Genet Dev Biol, 2000 Apr-Jun, 20(2), 107 - 8
Subsite specificity of divalent metal ions to glucosyltransferase; Devulapalle KS et al.; Glucosyltransferase from oral bacteria Streptococcus mutans is the most significant virulent factor in causing dental caries . The enzyme has two subsites . The binding specificity of divalent metal ions to glucosyl or fructosyl subsite was examined using multiple inhibition kinetics . The interaction factor "alpha" identifies whether the two subsites are exclusive or non-exclusive.

Conn Med, 2000 Oct, 64(10), 587 - 90
Culturing the throat to protect the heart: Dr . Milton Markowitz and the prevention of rheumatic fever; Murray T et al.; The purpose of this article is to briefly review how the understanding of rheumatic fever has evolved over the last 50 years . Particular emphasis is given to the identification of the Group A streptococcus as the causative agent of rheumatic fever and the use of antibiotics to treat and prevent rheumatic fever . Throughout his 50-year career, Dr . Milton Markowitz, former chairman of the department of pediatrics at the University of Connecticut School of Medicine, has been involved in these advances as an international expert on rheumatic fever prevention . Using archival materials, including some of the earliest literature descriptions of rheumatic fever, as well as an extended interview with Dr . Markowitz, the authors present a unique personal perspective both on the history of rheumatic fever and the context within which the scientific advances that have led to contemporary treatment and prevention strategies have evolved.

Microbes Infect, 2000 Oct, 2(12), 1425 - 30
The virulence of mixed infection with Streptococcus constellatus and Fusobacterium nucleatum in a murine orofacial infection model; Kuriyama T et al.; Orofacial infections are usually polymicrobial, and it is the microbial interactions of pathogenic species that cause tissue destruction . In this study, the microbial interaction between Streptococcus constellatus and Fusobacterium nucleatum was characterized using a murine orofacial infection model . A mixture of viable S . constellatus and F . nucleatum cells (both 2 x 10(8) CFU/mouse) was injected into the submandible; as a result, all of the test mice died . In contrast, none of the experimental animals monoinjected with either S . constellatus or F . nucleatum died (P<0.001), indicating that the synergism between the two resulted in the virulence . When a mixture of viable S . constellatus cells and a culture filtrate of F . nucleatum was tested, lethality and the bacterial cell count per lesion were significantly enhanced as compared with monoinjections (P<0.02) . However, the virulence of F . nucleatum was not enhanced by infection of a culture filtrate of S . constellatus . The enhancement of virulence was observed even when viable S . constellatus cells and the culture filtrate of F . nucleatum were injected at separate sites . Heat treatment of the culture filtrate of F . nucleatum did not affect the enhancement . These results indicate that a heat-stable substance(s) produced by F . nucleatum contributes to the microbial synergy of S . constellatus and F . nucleatum in orofacial infections.

Braz J Infect Dis, 1998 Apr, 2(2), 90 - 96
Characteristics of Isolates Streptococcus pneumoniae from Middle-Aged and Elderly Adults in Brazil: Capsular Serotypes and Antimicrobial Sensitivity to Invasive Infections; de Cunto Brandileone MC et al.; Pneumococcal infection cause frequent, serious problems among middle aged and elderly populations worldwide . Efforts to prevent mortality caused by pneumococci are based mainly on rapid diagnosis of the infection and appropriate antimicrobial therapy . Vaccination is considered to be the best approach to prevent the disease in at risk populations and the current 23-valent pneumococcal polysaccharide vaccine is recommended for people &gte;65 years of age . To evaluate the present antibiotic sensitivity patterns and the potential for vaccine use to prevent this disease in Brazil, 94 isolates of pneumococci from normally sterile body sites from patients &gte;50 years of age were analyzed for capsular serotypes and antimicrobial resistance . Among the total isolates, 7.4% (n = 7) of the isolates showed intermediate level resistance to penicillin (IR) . Among the IR isolates, 6 were also resistant to trimethoprim-sulfamethoxazole . All isolates were susceptible to cefotaxime, chloramphenicol, erythromycin, and vancomycin . Twenty-eight serotypes were identified and the serotypes included in the 23-valent vaccine accounted for 81.7% . By adding the cross-reacting serotypes, the percentage of vaccine preventable infections was 90.3% . Each of intermediate level resistant strains were among the 23 serotypes included in pneumococcal vaccine . This preliminary data on the distribution of serotype of S.pneumoniae among those &gte;50 years of age in Brazil, and the potential for increased antimicrobial resistance to penicillin and trimethoprim-sulfamethoxazole, support the use of pneumococcal vaccine in this population.

Pediatr Infect Dis J, 2000 Nov, 19(11), 1078 - 83
Bacteriology of histopathologically defined appendicitis in children; Rautio M et al.; BACKGROUND: Acute appendicitis is the most common surgical emergency in childhood . However, the pathogenesis and detailed microbiology are obscure . OBJECTIVE: To determine in detail the bacterial etiology of appendicitis in children in relation to the histologic tissue pathology . STUDY DESIGN: Tissue samples obtained at surgery from 41 children with suspected acute appendicitis were examined histologically and by culture for aerobic and anaerobic bacteria . The patients were analyzed according to histopathologic and clinical findings . RESULTS: Aerobic and anaerobic species were isolated from 40 of 41 (98%) samples; on average, 14.1 isolates per specimen (10.4 anaerobes and 3.7 aerobes) . Specimens from patients with gangrenous appendices yielded significantly higher numbers of anaerobic isolates per specimen than did specimens from patients with healthy appendices (11.7 vs . 7.7; P < 0.01) . Bacteria belonging to the Bacteroides fragilis group were the most frequently isolated anaerobic microorganisms (95%) . Other organisms frequently isolated in all histology groups were Peptostreptococcus micros (66%), Bilophila wadsworthia (63%), Fusobacterium nucleatum (44%), Eggerthella lenta (44%) and a hitherto undescribed bile-resistant, pigment-producing Gram-negative rod (41%) . Of the aerobes Escherichia coli (88%) and Streptococcus anginosus group (former Streptococcus "milleri" group) organisms (61%) were the most frequent findings . CONCLUSIONS: The shift from histologically normal toward gangrenous appendices was clearly associated with markedly elevated anaerobic bacterial counts in terms of species . The unusually high frequencies of B . wadsworthia (75%) and the hitherto undescribed bile-resistant, pigment-producing Gram-negative rod (56%) in gangrenous appendices represent unique and different findings from those reported in adults.

Can J Anaesth, 2000 Nov, 47(11), 1129 - 40
Severe group A streptococcal infection and streptococcal toxic shock syndrome; Baxter F et al.; PURPOSE: To review the literature on group A streptococcal toxic shock syndrome, (STSS) . DATA SOURCE: Medline and EMBASE searches were conducted using the key words group A streptococcal toxic shock syndrome, alone and in combination with anesthesia; and septic shock, combined with anesthesia . Medline was also searched using key words intravenous immunoglobulin, (IVIG) and group A streptococcus, (GAS); and group A streptococcus and antibiotic therapy . Other references were included in this review if they addressed the history, microbiology, pathophysiology, incidence, mortality, presentation and management of invasive GAS infections . Relevant references from the papers reviewed were also considered . Articles on the foregoing topics were included regardless of study design . Non-English language studies were excluded . Literature on the efficacy of IVIG and optimal antibiotic therapy was specifically searched . PRINCIPAL FINDINGS: Reports of invasive GAS infections have recently increased . Invasive GAS infection is associated with a toxic shock syndrome, (STSS), in 8-14% of cases . The STSS characteristically results in shock and multi-organ failure soon after the onset of symptoms, and is associated with a mortality of 33-81% . Many of these patients will require extensive soft tissue debridement or amputation in the operating room, on an emergency basis . The extent of tissue debridement required is often underestimated before skin incision . CONCLUSIONS: Management of STSS requires volume resuscitation, vasopressor/inotrope infusion, antibiotic therapy and supportive care in an intensive care unit, usually including mechanical ventilation . Intravenous immunoglobulin infusion has been recommended . Further studies are needed to define the role of IVIG in STSS management and to determine optimal anesthetic management of patients with septic shock.

Clin Infect Dis, 2000 Dec, 31(6), 1373 - 9 Epub 2000 Nov 29.
Antibiotic susceptibility and serotype distribution of Streptococcus pneumoniae causing meningitis in Italy, 1997-1999; Pantosti A et al.; Because few data are available in Italy regarding antimicrobial susceptibility and serotype distribution of invasive Streptococcus pneumoniae strains, meningeal isolates collected at Italian hospitals during the years 1997-1999 were studied . The 12 most common serogroups, representing > 85% of the isolates, were 14, 23, 6, 4, 3, 9, 19, 8, 1, 12, 18, and 7 (in order of frequency) . The serogroups identified in children < 5 years old were more limited in number: 80% are included in the 7-valent conjugate vaccines . Penicillin resistance was observed in 14 (9.5%) of 148 strains and increased from 5% in the first part of the study to 13% in the last part . Only 2 strains were fully penicillin resistant, and these belonged to serotype 9V . Thirty percent of the strains, mostly belonging to serogroups 14 or 6 and carrying either the ermB or the mef genes, were resistant to erythromycin.

South Med J, 2000 Nov, 93(11), 1096 - 8
Necrotizing fasciitis and toxic shock-like syndrome caused by group B streptococcus; Holmstrom B et al.; A recent increase in reports of necrotizing fasciitis resulting from group B streptococcus has alerted physicians to a possible concomitant increase of toxic shock-like syndrome . We report the second case of group B streptococcus causing necrotizing fasciitis and toxic shock-like syndrome . A black woman, aged 52 years, with newly diagnosed diabetes mellitus had necrotizing fasciitis type II of the left groin . Hypotension, elevated bilirubin and liver enzymes, and adult respiratory distress syndrome rapidly developed . Because group B streptococcus was isolated from a normally sterile site, the patient's condition met the criteria for toxic shock-like syndrome . Extensive surgical debridement, hyperbaric oxygen therapy, and intravenous antibiotic therapy (including clindamycin) were required for complete recovery . The antitoxin effects of hyperbaric oxygen therapy and clindamycin should be further investigated for the treatment of such patients.

South Med J, 2000 Nov, 93(11), 1078 - 80
Prevalence of penicillin-nonsusceptible pneumococcal bacteremia in a Staten Island community hospital; Perrone C et al.; BACKGROUND: Although the first reports of infection due to penicillin-nonsusceptible Streptococcus pneumoniae in the United States were in children, these strains have circulated widely in recent years, with the prevalence increasing dramatically among the elderly . Regional surveillance of pneumococcal susceptibility profiles may assist clinicians in management decisions, increase awareness of this microbial threat, and target potential areas of intervention . METHODS: As part of ongoing surveillance, we surveyed single-patient pneumococcal blood isolates in our 440-bed Staten Island community teaching hospital from June 1, 1996, through May 31, 1998 . RESULTS: Overall, of 47 single-patient isolates, 16 (35%) were penicillin nonsusceptible . Of 35 isolates from adults, 15 (44%) were nonsusceptible, compared with 1 of 12 (8%) from children . Seven of the nonsusceptible isolates (44%) were from persons > or = 65 years old and represented 47% of the isolates from this age group . CONCLUSIONS: Community-acquired penicillin-nonsusceptible pneumococcal bacteremia is not simply a pediatric problem, but also a threat to the elderly.

Int J Dermatol, 2000 Oct, 39(10), 779 - 83
A century of the synthesis of dapsone: its anti-infective capacity now and then; Wolf R et al.; BACKGROUND: Although dapsone was first synthesized in 1908, a quarter of a century was to pass before it was used in the treatment of bacterial infections . Dapsone was, however, too toxic for humans (because of the excess dosage which was administered at that time) and was thus considered to be of no value in the treatment of common bacterial infections . Since the early 1950s, dapsone has been recognized as being uniquely effective against a number of noninfectious, inflammatory diseases and, today, this is its main indication . Thus, the reason why dapsone was first introduced into medicine, namely the treatment of bacterial infections, has been set aside and its main current applications are the treatment of noninfectious, inflammatory, autoimmune, and bullous diseases . OBJECTIVE: To study the anti-infective capacity of dapsone against common bacterial infections . As many patients who receive dapsone for the treatment of noninfectious, inflammatory diseases have a concomitant bacterial infection or a superinfection of their skin disease, we thought that, if dapsone proved to be effective against common bacterial infections, it may obviate the need for an additional antimicrobial drug in these patients . METHODS: Three bacterial ATCC> strains (Streptococcus pyogenes, Staphylococcus aureus, and Escherichia coli) were tested by a macrodilution minimal inhibitory concentration (MIC) test for dapsone . Dapsone concentrations were between 0.06 and 1125 microg/mL . RESULTS: Even the highest concentration of dapsone of 1125 microg/mL did not inhibit bacterial growth . CONCLUSIONS: Our results indicate that dapsone has no antibacterial effects whatsoever . Even at very high concentrations, it does not suppress the growth of most susceptible strains of bacteria . The story of dapsone (i.e . the long time that elapsed between its synthesis to its use for the chemotherapy of infectious diseases) will not repeat itself this time.

Indian J Med Res, 2000 Sep, 112, 100 - 3
Throat carriage of pneumococci in healthy school children in the Union Territory of Pondicherry; Kanungo R et al.; BACKGROUND & OBJECTIVES: Colonisation of Streptococcus pneumoniae in the throat is common among children the world over . Little is known about the relationship of nasopharyngeal carriage and invasive disease or the way it spreads within the households and close confines . There is a paucity of data on the colonization of Strep . pneumoniae in the throat of healthy children in India . To determine the prevalence of pneumococcal carriage in school children of urban and rural Pondicherry, a study was undertaken . METHODS: Throat swabs of healthy school-going children between 5-10 yr of age were examined for pneumococcal carriage, by standard bacteriological techniques . RESULTS: A prevalence rate of 24.3 per cent was noted . There was no difference in the carriage rate among the rural children when compared to urban children . No age, sex or geographical predilection of pneumococcal carriage was noted . A statistically significant seasonal variation, however, was seen . Carriage rate increased during the colder months and was found to be the highest in the months of March and November . INTERPRETATION & CONCLUSIONS: Strep . pneumoniae circulates in the community among healthy children . Carriage rate is influenced by seasonal variation.

Curr Treat Options Cardiovasc Med, 2000 Dec, 2(6), 481 - 488
Endocarditis in the Pediatric Population; Stock JH et al.; Endocarditis is a rare, but potentially fatal process in children . Patients with congenital heart disease compose the majority of patients with endocarditis . Neonates and children with central venous catheters are an increasingly frequent group of patients diagnose with this disease . Rheumatic fever predisposing to endocarditis is unusual . Streptococcus viridans and Staphylococcus aureus are the most pervasive organisms associated with endocarditis, though others are becoming more frequent . Blood cultures should be obtained in febrile children with congenital heart disease before the administration of antibiotics . Echocardiography is useful in children with known endocarditis, and in children in whom there is a high level of clinical suspicion for endocarditis . Echocardiography is a poor screening tool for patients without clinical or bacteriologic evidence for endocarditis . Endocarditis prophylaxis for children with congenital heart disease (excluding a secundum atrial septal defect) before appropriate procedures is recommended.

J Biol Chem, 2001 Mar 16, 276(11), 8197 - 204 Epub 2000 Nov 28.
Streptococcal IgA-binding proteins bind in the Calpha 2-Calpha 3 interdomain region and inhibit binding of IgA to human CD89; Pleass RJ et al.; Certain pathogenic bacteria express surface proteins that bind to the Fc part of human IgA or IgG . These bacterial proteins are important as immunochemical tools and model systems, but their biological function is still unclear . Here, we describe studies of three streptococcal proteins that bind IgA: the Sir22 and Arp4 proteins of Streptococcus pyogenes and the unrelated beta protein of group B streptococcus . Analysis of IgA domain swap and point mutants indicated that two loops at the Calpha2/Calpha3 domain interface are critical for binding of the streptococcal proteins . This region is also used in binding the human IgA receptor CD89, an important mediator of IgA effector function . In agreement with this finding, the three IgA-binding proteins and a 50-residue IgA-binding peptide derived from Sir22 blocked the ability of IgA to bind CD89 . Further, the Arp4 protein inhibited the ability of IgA to trigger a neutrophil respiratory burst via CD89 . Thus, we have identified residues on IgA-Fc that play a key role in binding of different streptococcal IgA-binding proteins, and we have identified a mechanism by which a bacterial IgA-binding protein may interfere with IgA effector function.

Curr Infect Dis Rep, 2000 Oct, 2(5), 399 - 408
Impact of Antimicrobial Resistance on the Treatment of Invasive Pneumococcal Infections; Rocha P et al.; Infectious diseases, such as pneumococcal pneumonia, which were almost invariably lethal in the pre-antibiotic era, caused radically less mortality with the advent of antimicrobial chemotherapy . However, the use, misuse, and abuse of these agents have led to the emergence of antimicrobial resistance . In the past, pneumococci were all exquisitely sensitive to penicillin G . By the late 1960s, penicillin-nonsusceptible Streptococcus pneumoniae (PNSP) was being described . Since then, this problem has achieved epidemic proportions in many areas of the world, including the United States . Many experts no longer consider penicillin an acceptable therapy for patients suspected of having invasive pneumococcal disease, especially if the central nervous system (CNS) is involved . Recommendations for therapy are based on theoretical concerns, in vitro susceptibility testing, animal data, and a few, scattered reports of penicillin failure in patients with invasive disease.

Curr Infect Dis Rep, 2000 Jun, 2(3), 207 - 214
Gram-Positive Pneumonia; Osiyemi O et al.; Gram-positive pneumonia is a leading cause of morbidity and mortality throughout the world . Of the gram-positive pathogens that cause pneumonia, Streptococcus pneumoniae and Staphylococcus aureus are the most common . The diagnosis of gram-positive pneumonia remains less than satisfactory, and newer diagnostic techniques such as antibody- and polymerase chain reaction-based antigen detection have yet to prove themselves . Drug resistance among gram-positive organisms is now endemic throughout the world and remains a serious therapeutic problem despite the availability of new antimicrobials . Efforts to control the spread of resistant strains include, in the case of S . aureus, stringent isolation policies and topical treatment to reduce carriage and, for S . pneumoniae, increased use of available vaccines and the develop- ment of more immunogenic vaccines.

Curr Infect Dis Rep, 1999 Dec, 1(5), 470 - 479
New Fluoroquinolones: Real and Potential Roles; Ball P; The second-generation fluoroquinolones have enjoyed successful clinical use for more than 10 years in many countries, and they have a valued and proven record of safety and efficacy . However, deficiencies with respect to gram-positive and anaerobic organisms limit the use of these agents in respiratory, intra-abdominal, and pelvic infections . New, third-generation agents with dramatically increased activity against gram-positive and anaerobic bacteria--notably, Streptococcus pneumoniae and Bacteroides fragilis--have shown high rates of efficacy in pneumonia, bronchitis, and surgical and gynecologic infections . Although most of these new drugs produce similar clinical results, adverse reaction profiles differ and may influence therapeutic choices.

Curr Infect Dis Rep, 1999 Dec, 1(5), 458 - 463
Ribosomal Resistance: Emerging Problems and Potential Solutions; Zhanel GG et al.; Many systemic antibiotics use ribosomal inhibition to suppress the replication of bacteria . Current research suggests that resistance to macrolide, lincosamide, and streptogramin B (MLS(B)) antibiotics is emerging among clinical isolates of Streptococcus pyogenes and Streptococcus pneumoniae . Erythromycin methylases, encoded by erm genes, modify an essential adenine residue in 23S rRNA and confer cross-resistance to MLS(B) antibiotics . More recently, macrolide efflux (mef) genes were identified in isolates of S . pyogenes and S . pneumoniae that show resistance to 14- and 15-membered macrolides (M phenotype) . Resistance to MLSB has been associated with the increased use of erythromycin, and the recent emergence of the M phenotype has coincided with the marketing of newer macrolides . However, despite increasing macrolide resistance among clinical isolates of S . pneumoniae, convincing data on treatment failures directly attributable to MLS(B) or M phenotypes are limited . Possible solutions to emerging MLS(B) and M phenotype resistance include the introduction of alternative antibiotics, the more prudent use of antibiotics, combination therapy, molecular diagnostics, enhanced understanding of pharmacodynamic variables, and redefined resistance breakpoints.

Curr Infect Dis Rep, 1999 Apr, 1(1), 57 - 64
Pneumococcal Pneumonia; Mufson MA; Pneumococcal pneumonia accounts for about one-sixth to two-thirds of all cases of community-acquired pneumonia . Its high frequency of occurrence worldwide and the high number of deaths associated with it--especially with bacteremic (invasive) disease--mark its importance . Invasive disease is associated with case-fatality rates of 15% to 25% among elderly adults . Penicillin-resistant Streptococcus pneumoniae (PRSP) first appeared in the 1970s, and its increased incidence in the late 1980s signaled its emerging importance . In individual patients in whom PRSP infection is suspected, the clinician must follow guidelines for empiric antibiotic therapy for community-acquired pneumonia until microbiological test results are known . When a diagnosis of pneumococcal pneumonia is established, the clinician should change to a regimen that targets the pneumococcus . Adults at highest risk for death from pneumococcal pneumonia include immunocompetent persons with underlying chronic diseases, immunocompromised persons, elderly persons, and unvaccinated residents of nursing homes and other chronic care facilities . Safe and effective, polyvalent polysaccharide pneumococcal vaccine should be used in persons 2 years of age and older who are at increased risk for serious pneumococcal pneumonia and in all persons 65 years of age and older.

Curr Infect Dis Rep, 1999 Apr, 1(1), 49 - 56
Community-Acquired Pneumonia: An Overview; Kanno MB et al.; Community-acquired pneumonia (CAP) accounts for a significant number of hospitalizations and outpatient visits, as well as substantial health care expenditures . CAP is particularly common among the elderly who account for more than 90% of deaths due to pneumonia . Streptococcus pneumoniae is believed to be the most common microbial etiology of CAP, but recent studies suggest that the atypical pathogens may be more common than previously thought, particularly among ambulatory patients . Recent studies have provided data regarding risk of mortality and process of care and outcomes . Increasing resistance among strains of S . pneumoniae has impacted the approach to the empiric therapy of CAP . The Infectious Diseases Society of America published guidelines for the evaluation and management of CAP this past year . Pathogen-specific therapy guided by the results of sputum gram stain and culture is emphasized . Despite enthusiasm for practice guidelines and clinical pathways, there remains insufficient published data to determine their impact on quality and cost of care in patients with CAP.

Curr Infect Dis Rep, 1999 Apr, 1(1), 13 - 21
Streptococcus pneumoniae: Activity of Newer Agents Against Penicillin-Resistant Strains; Jacobs MR et al.; Strains of pneumococci resistant to antimicrobial agents have been reported on all continents . In 1997, more than 50% of strains in the United States were not susceptible to penicillin, and 30% were resistant to macrolides . In addition, many strains are resistant to multiple agents, including beta-lactams, macrolides, clindamycin, chloramphenicol, tetracyclines, and trimethoprim-sulfamethoxazole . Although resistance to beta-lactams in nonmeningeal infections can usually be overcome by parenteral administration, clinically significant resistance is an important limitation in meningitis and with oral administration of beta-lactams . Decisions about treatment of pneumococcal infection are based on the site of infection, the degree of resistance to penicillin G, the presence of resistance to other agents, the severity of disease, the presence of underlying conditions, and the dose and route of administration of antimicrobial agents . The application of pharmacokinetic and pharmacodynamic variables to pneumococci has greatly improved the interpretation of susceptibility data and the development of clinically relevant breakpoints.

Jpn J Ophthalmol . 2000 Nov 1;44(6):692.
Molecular Biological Analysis of Retinal and Streptococcal Heat-shock Protein 60; Tanaka T et al.; Purpose: The observation of recurrent oral aphtha preceding ocular inflammation in patients with Behcet's disease suggests a role of oral Streptococcus in the etiology of this disease . Heat-treated Streptoccus antigen can induce ocular inflammation or systemic symptoms in Behcet's disease patients . Furthermore, the presence of an autoantibody against retinal heat-shock protein 60 (HSP 60) has been detected in the sera of these patients . Injection of extracted retinal HSP 60 also induces experimental uveitis in treated rats . The characteristics of retinal HSP 60 and HSP 60 from S . pyogenes were evaluated using a molecular biological approach.Methods: The gene encoding HSP 60 was isolated by polymerase chain reaction (PCR) from a bovine retinal cDNA library and from S . pyogenes DNA . The DNA sequence of the HSP 60 coding region was determined, and the amino acid sequences of the encoded proteins were predicted from the DNA sequence . The synthetic peptide (50 nmol) was emulsified with Freund's complete adjuvant and injected into rats.Results: Comparison of the amino acid sequences of Streptococcal and bovine retinal HSP 60 revealed about 200 residue regions with 47% homology . Experimental uveitis was mainly induced in rats inoculated with retinal HSP 60-derived peptide or Streptococcus HSP 60-derived peptide that is equivalent to residues 245-259 of human HSP 65.Discussion: The retinal and Streptococcal HSP 60 amoino acid composition is thought to be important and useful for investigating the mechanisms involved in the induction of Behcet's disease

Equine Vet J, 2000 Nov, 32(6), 527 - 32
Elimination of guttural pouch infection and inflammation in asymptomatic carriers of Streptococcus equi; Verheyen K et al.; Three protracted outbreaks of strangles were investigated using endoscopic examination and a total of 14 asymptomatic carriers of Streptococcus equi were identified of which 13 showed evidence of carriage in the guttural pouch . Treatment was initiated to eliminate S . equi colonisation since these animals posed an infectious risk to susceptible horses . Two further horses were referred to us with severe guttural pouch pathology and from which S . equi was cultured, and treatment of these cases is also described . Treatment in the first instance was directed towards removal of gross guttural pouch pathology as seen on endoscopic examination . This was done with a combination of irrigation of the pouch with moderate to large amounts of saline, suction of fluid material and endoscopic manipulation of chondroids . Subsequently, antibiotic treatment was used to eliminate S . equi infection . All animals received systemic antibiotics, in some cases combined with topical antimicrobial treatment . Treatment was generally regarded as successful when the guttural pouches appeared normal and S . equi was not detected in nasopharangeal swabs and pouch lavages on 3 consecutive occasions . Successful treatment of one carrier required surgical intervention due to occlusion of both guttural pouch pharyngeal openings . Fourteen of 15 carriers were successfully treated by endoscopic removal of inflammatory material and antibiotic treatment, without surgical intervention . Five carriers originally given potentiated sulphonamide (33%) required further therapy with penicillin or ceftiofur, administered both systemically and topically, before S . equi infection and associated inflammation of the guttural pouches were eliminated.

Equine Vet J, 2000 Nov, 32(6), 515 - 26
Control of strangles outbreaks by isolation of guttural pouch carriers identified using PCR and culture of Streptococcus equi; Newton JR et al.; Previous use of repeated nasopharyngeal swabbing and culture of Streptococcus equi showed that healthy carriers developed in more than 50% of 'strangles' outbreaks . The guttural pouches were the only detectable site of S . equi colonisation on endoscopic examination of horses during one of these outbreaks and S . equi was sometimes not detected by culture of nasopharyngeal swabs from carriers for up to 2 or 3 months before nasal shedding resumed sporadically . A more sensitive way of detecting S . equi on swabs from established guttural pouch carriers was therefore required . Conveniently selected 'strangles' outbreaks were investigated in detail using endoscopy, in order to develop and assess a suitable polymerase chain reaction (PCR) test . We report here 3 protracted 'strangles' outbreaks on different kinds of establishments in which between 29 and 52% of sampled horses were infected as detected by culture and/or PCR . Of the infected horses, between 9 and 44% were identified as carrying S . equi after clinical signs had disappeared and the predominant site of carriage was the guttural pouch . Prolonged carriage of S . equi, which lasted up to 8 months, did not cease spontaneously before treatment was initiated to eliminate the infections . The detection and isolation of the carriers, in conjunction with strict hygiene measures, apparently resulted in the control of the outbreaks and allowed the premises to return to normal activity . Comparing PCR and culture, many more swabs were found to be positive using PCR (56 vs . 30% of 61 swabs) . Similar results were obtained for guttural pouch samples from 12 established carriers (PCR 76% and culture 59%) . These results from repeated samples from relatively few animals need confirming using more long-term carriers . PCR can also detect dead organisms and is, therefore, liable to yield false positive results . Despite this drawback, it is argued that PCR provides a potentially useful adjunct to culture of nasopharyngeal swabs in the detection of asymptomatic carriers of S . equi following outbreaks of 'strangles'.

Vet Rec, 2000 Oct 21, 147(17), 480 - 4
Endoscopic and bacteriological findings in a chronic outbreak of strangles; Fintl C et al.; Recently there has been increased awareness of the role of the carrier state in propagating Streptococcus equi var equi (S equi) infections (strangles), although the anatomical location of the organisms in chronic carriers has not been consistently established . This case report describes a chronic strangles outbreak in a riding school, that was monitored over six months by repeated clinical and endoscopic guttural pouch examinations . All asymptomatic horses that had positive S equi cultures on nasal swabs or guttural pouch lavages were found to have lesions in their guttural pouches . These lesions included empyema, chondroids and previously undescribed chronic discharging lesions on the floor of the medical compartment of the guttural pouches . These observations further support previous studies indicating the importance of investigating the guttural pouches in horses suspected to be asymptomatic carriers of this organism.

Caries Res, 2000 Nov-Dec, 34(6), 498 - 501
Antimicrobial effect of a novel ozone- generating device on micro-organisms associated with primary root carious lesions in vitro; Baysan A et al.; The aims of this present study were (1) to assess the antimicrobial effect of ozone from a novel ozone-generating device (Heolozone, USA) {0.052% (v/v) in air delivered at a rate of 13.33 ml.s(-1)} on primary root carious lesions (PRCLs) and (2) to evaluate the efficacy of ozone specifically on Streptococcus mutans and Streptococcus sobrinus . In study 1, 40 soft PRCLs from freshly extracted teeth were randomly divided into two groups to test the antimicrobial effect on PRCLs from exposure to ozonated water for either 10 or 20 s . Half of a lesion was removed using a sterile excavator . Subsequently, the remaining lesion was exposed to the ozonised water for a period of either 10 or 20 s (corresponding to 0 . 069 or 0.138 ml of ozone, respectively) . Using paired Student t tests, a significant (p<0.001) reduction (mean +/- SE) was observed in the ozone-treated groups with either a 10-second (log(10) 3.57+/-0.37) or 20-second (log(10) 3.77+/-0.42) ozone application compared with the control groups (log(10) 5.91+/-0.15 and log(10) 6.18+/-0.21, respectively) . In study 2, 40 sterile saliva-coated glass beads were randomly divided into two groups for each micro-organism . One glass bead was put into each bijou bottle with 3 ml of Todd-Hewitt broth . S . mutans and S . sobrinus were inoculated anaerobically overnight . Each glass bead was then washed with 2 ml of phosphate-buffered saline . Immediately, 10 s of ozone gas was applied to each glass bead in the test groups . There was a significant (p<0.0001) reduction (mean +/- SE) in ozone-treated samples for S . mutans (log(10) 1.01+/-0.27) and S . sobrinus (log(10) 1.09+/-0.36) compared with the control samples (log(10) 3.93+/-0.07 and log(10) 4.61+/-0.13, respectively) . This treatment regime is an effective, quick, conservative and simple method to kill micro-organisms in PRCLs . Ozone gas application for a period of 10 s was also capable of reducing the numbers of S . mutans and S . sobrinus on saliva-coated glass beads in vitro.

Microbiol Immunol, 2000, 44(9), 755 - 64
Nucleotide sequencing and transcriptional analysis of two tandem genes encoding glucosyltransferase (water-soluble-glucan synthetase) in Streptococcus cricetus HS-6; Inoue M et al.; Two tandem genes encoding glucosyltransferase synthesizing water-soluble glucan (GTF-S) were cloned from the lambda gene library of Streptococcus cricetus HS-6 (serotype a) using anti-GTF-S antibody, and the nucleotide sequences were analyzed . The two genes (ORF1 and ORF2) were identified as streptococcal glucosyltransferases based on the following evidence: {1} the deduced amino acid sequences of their products have an active site for catalytic action and C-terminal repeated units for dextran binding, and {2} a homology search revealed that the ORF1 and ORF2 products are homologous to the GtfS protein (77.4%) of S . downei Mfe28 and GtfT protein (83.8%) of S . sobrinus OMZ176, respectively, which are both known to have GTF-S activity . Therefore, ORF1 and ORF2 might be designated gtfS and gtfT of S . cricetus, respectively . A Northern blotting and RNase protection assay suggested that the gtfS and gtfT of S . cricetus are transcribed as a single bicistronic mRNA as well as separate monocistronic mRNAs . Primer extension analysis indicated multiple transcriptional start points for each gene.

Clin Exp Immunol, 2000 Nov, 122(2), 207 - 11
Treatment with a monocolonal antibody to IL-8 attenuates the pleocytosis in experimental pneumococcal meningitis in rabbits when given intravenously, but not intracisternally; Ostergaard C et al.; The role of interleukin (IL)-8 as mediator in the recruitment of leucocytes into the CSF was investigated during experimental pneumococcal meningitis . Rabbits were inoculated intracisternally with approximately 10(6) CFU Streptococcus pneumoniae, and treated (i) intravenously with 5 mg of a monoclonal antibody to IL-8 (n = 7) or 5 mg of an isotype control antibody (n = 6); (ii) intracisternally with anti-IL-8, 100 microg (n = 5), 10 microg (n = 4), 1 microg (n = 4), 0.1 microg (n = 2) . Ten rabbits served as untreated control group . Intravenous treatment with anti-IL-8 attenuated the pleocytosis significantly compared to untreated rabbits (P < 0.04) or rabbits treated with an isotype control antibody (P < 0.02) . In contrast, intracisternal treatment with anti-IL-8 failed to attenuate the pleocytosis (P > 0.05) . These results show, that IL-8 plays an important role in the recruitment of leucocytes during experimental pneumococcal meningitis, and that the functional activity of IL-8 in this process appears to be on the bloodstream side of the microvascular endothelium of the brain.

Int J Antimicrob Agents, 2000 Nov, 16(3), 335 - 9
Emerging and reemerging pathogens; Gomez-Lus R et al.; From 1973 to 1995, 29 new and reemerging pathogenic microbes were recognized . However, in discussions about emerging infectious diseases, the focus is often on the clinical effects of the host-parasite relationship, rather than the examination of the biology of the pathogen . Many of what we refer to as emerging diseases are characterized better as 'diseases of human progress' . Thus, the aerosolization of water has played an important role in the emergence of Legionella pneumophila infections . New diseases are superimposed on endemic diseases such as diarrhoeal diseases, malaria and tuberculosis . In addition, many pathogens are becoming increasingly resistant to standard antimicrobial drugs, making treatment difficult and in some cases impossible . We summarize our experience on emerging parasitic diseases (primary amoebic meningoencephalitis, respiratory cryptosporidiosis, and diplogonoporiasis), and selected problems of bacterial resistance (MDR tuberculosis caused by Mycobacterium bovis and macrolide-resistance mechanisms of Streptococcus pneumoniae and S . pyogenes).

Int J Antimicrob Agents, 2000 Nov, 16(3), 219 - 24
Mediterranean clone of penicillin-susceptible, multidrug-resistant serotype 6B Streptococcus pneumoniae in Greece, Italy and Israel; Syrogiannopoulos GA et al.; In 1996, 19 isolates of serotype 6B Streptococcus pneumoniae with a unique resistance pattern were found in carriers attending daycare centres in Patras, Southwestern Greece . These isolates were penicillin susceptible but resistant to chloramphenicol, tetracycline, erythromycin, clindamycin and trimethoprim-sulphamethoxazole . Subsequently, isolates with the same characteristics were found in 23 additional carriers in central and southern Greece in 1997-98 as well as in 19 carriers in central Italy in 1997, and in seven carriers in southern Israel in 1998 . Carriers were all children under 6 years of age, attending daycare centres or outpatient hospital visits . The relatedness of the isolates was determined on representative isolates from the three countries by pulsed-field gel electrophoresis of SmaI digests of chromosomal DNA . Most Greek isolates were identical to each other, while isolates from Italy and Israel showed one to three band differences, with all isolates being closely related to each other as well as to the isolates from Greece . We have therefore documented the presence of this unique clone of S . pneumoniae in these three countries and have named this the 'Mediterranean' clone . While isolates appear to have a common origin, their source and direction of spread are unknown . However, isolates from Italy showed the most diversity, suggesting that this clone had been present in that country for a longer period than it had been in Greece.

AJR Am J Roentgenol, 2000 Dec, 175(6), 1533 - 6
Pneumococcal pneumonia in patients requiring hospitalization: effects of bacteremia and HIV seropositivity on radiographic appearance; Shah RM et al.; OBJECTIVE: Our purpose was to establish the incidence of lobar versus bronchopneumonia patterns in patients hospitalized for pneumococcal pneumonia and to determine whether bacteremia or HIV status affects the radiographic appearance . MATERIALS AND METHODS: Eighty-one patients with community-acquired pneumococcal pneumonia and positive findings on admission chest radiographs were selected from a group of 105 patients with positive findings for Streptococcus pneumoniae from sputum or blood cultures . Patients less than 16 years old and those with nosocomial pneumonia, aspiration pneumonia, or coexistent lung disease were excluded . The dominant pattern was classified as lobar or lobular bronchopneumonia by two radiologists who were unaware of results from blood cultures and HIV testing . RESULTS: Forty-three females and 38 males, with a mean age of 48 years (age range, 16-92 years), required admission for pneumococcal pneumonia . Fifty-nine (73%) of the 81 patients were bacteremic and 20 (25%) of the 81 patients were HIV-positive, including 14 patients (17%) who were both bacteremic and HIV-positive . Among all the patients, focal lobar consolidation was the most common pattern, observed in 48% . Multifocal lobar consolidation was the next most frequent pattern, occurring in 33% . Multifocal and focal bronchopneumonia patterns were seen in 16% and 2% of the patients, respectively . Overall, multifocal consolidation occurred in 49% . The dominant radiographic pattern and incidence of multicentric disease were not affected by HIV seropositivity (p = 0 . 61) or bacteremia (p = 0.17) . CONCLUSION: Lobar consolidation, involving single or multiple lobes, is the most common radiographic pattern of community-acquired pneumococcal pneumonia in patients requiring hospitalization . The pattern of consolidation is not influenced by bacteremia or HIV status.

Ann Otol Rhinol Laryngol, 2000 Nov, 109(11), 1021 - 7
Effect of adenovirus type 1 and influenza A virus on Streptococcus pneumoniae nasopharyngeal colonization and otitis media in the chinchilla; Tong HH et al.; Considerable evidence has implicated respiratory tract virus potentiation of bacterial adherence, colonization, and superinfection as a significant factor contributing to the pathogenesis of otitis media (OM) . Influenza A and B viruses, adenovirus, and respiratory syncytial virus are the primary respiratory tract viruses associated with this disease . Investigations have established a dramatic increase in the development of experimental OM in chinchillas co-inoculated with influenza A virus and Streptococcus pneumoniae (Spn) . The mechanism underlying this phenomenon was suggested to involve, in part, viral compromise of eustachian tube mucosal integrity and function . This study was designed to assess and compare the effect of adenovirus and influenza A virus infection on adherence, the kinetics of colonization, and invasion of the middle ear by Spn in the chinchilla model of OM . Cohorts were inoculated intranasally with adenovirus type 1 or influenza A virus, and then inoculated intranasally 7 days later with Spn 6A . All cohorts were observed over a 14-day period after challenge with Spn, and the incidence and severity of OM were assessed by several methods, including culture of the nasopharynx and middle ear effusions . The data indicated that influenza A virus promotes a significant increase in nasopharyngeal colonization by Spn, an increased incidence and severity of OM, and a sustained presence of Spn in the effusions . Adenovirus infection, however, did not enhance colonization by Spn or result in an increased incidence or severity of OM.

Infect Control Hosp Epidemiol, 2000 Nov, 21(11), 711 - 7
Streptococcus pneumoniae serotype 4 outbreak in a home for the aged: report and review of recent outbreaks; Gleich S et al.; OBJECTIVE: To describe a pneumonia outbreak caused by Streptococcus pneumoniae among residents of a home for the aged and to review contemporary pneumococcal outbreaks . DESIGN: Epidemiological investigation . METHODS: S pneumoniae isolates were serotyped and analyzed by pulsed-field gel electrophoresis . Paired sera were tested for antibodies to pneumococcal surface adhesin A protein (PsaA, a 37-kDa cell-wall protein) . Pneumococcal outbreaks reported in the last decade in English were reviewed . RESULTS: Pneumonia developed in 18 of 200 residents . In 11 (61%), a pneumococcal etiology was demonstrated . S pneumoniae, serotype 4, was isolated from the blood cultures of 3 patients; all isolates were indistinguishable by pulsed-field gel electrophoresis . Pneumococcal involvement was established in 2 by sputum culture and latex agglutination of parapneumonic fluid and in 6 others by a twofold rise in optical density of serum antibody reactive to PsaA . Pneumococcal immunization had not previously been received by any patient; mortality was 22% . No additional cases were noted following administration of pneumococcal vaccine and antibiotic prophylaxis with penicillin or erythromycin . Twenty-six outbreaks of invasive pneumococcal disease since 1990 were reviewed . Twelve occurred in the United States, and serotypes 23F, 14, and 4 accounted for 8 (67%) of 12 outbreaks . All confirmed serotypes in US outbreaks are included in the 23-valent vaccine . More than one half of pneumococcal outbreaks worldwide involved elderly persons in hospitals or long-term-care facilities . CONCLUSIONS: A pneumococcal pneumonia outbreak occurred among unvaccinated residents of a residential facility for the aged . Institutionalized elderly persons are at risk of outbreaks of pneumococcal disease and should be vaccinated.

Laryngorhinootologie, 2000 Oct, 79(10), 609 - 15
{Intracranial otogenic complications: inspite of therapeutic progress still a serious problem}; Kaftan H et al.; BACKGROUND: High-imaging modalities, antibiotics and improved microsurgical procedures have decreased morbidity and mortality of intracranial otogenic complications in the western hemisphere nowadays . However, they do occur and the resulting mortality is still about 10% . PATIENTS AND RESULTS: We report our experiences with 22 patients, who were treated for intracranial otogenic complications (15 x meningitis, 5 x brain abscess, 1 x subdural empyema, 1 x Gradenigo's syndrome) due to cerebrospinal fluid leak in 2 patients, chronic otitis in 9 patients (5 with cholesteatoma), acute otitis media in 11 patients, between 1981 and September 1999 . Five patients with acute otitis media have had predisposing anatomic pathology: 1 x inner ear malformation, 1 x status after duraplasty, 2 x dura-brain-prolapse due to temporal bone fractures and 1 x dura-brain-prolapse after antrotomy . Streptococcus pneumoniae was a common cause of intracranial-complicating acute otitis media (64%) . Residual neurologic impairment was noted in 3 patients at the time of discharge . 3 patients (13.6%) died due to the otogenic intracranial complication . CONCLUSION: Otogenic intracranial complications are potentially life threatening conditions . Early diagnosis is essential to allow appropriate antimicrobial and surgical treatment . The necessity of close cooperation between otorhinolaryngologist, pediatrician, neurologist, radiologist and neurosurgeon is stressed.

Protein Expr Purif, 2000 Dec, 20(3), 379 - 88
Production, characterization, and crystallization of truncated forms of pneumococcal surface protein A from Escherichia coli; Lamani E et al.; Streptococcus pneumoniae is a major bacterial pathogen that causes diseases such as pneumonia and meningitis in humans . One of the antigens of this organism is pneumococcal surface protein A (PspA) . PspA is a virulence factor of the bacteria that has been shown to protect mice against pneumococcal infection . Among several domains of the protein, the amino-terminal part of PspA has been found to be a functional module which is essential for full pneumococcal infectivity . In order to investigate the properties of this protein, several internal fragments of the pspA gene were amplified from S . pneumoniae strain Rxl using the polymerase chain reaction (PCR) . The fragments were then cloned and expressed in Escherichia coli in a soluble form using the T7 RNA polymerase pET15b and pET21a vector systems . The size of these fragments ranges from 24 to 32 kDa corresponding to amino acids 67-272 (PspA-206), 1-236 (PspA-236), and 1-272 (PspA-272) . The fragments were purified to homogeneity using nickel chelating affinity, size exclusion, and anion-exchange chromatographic methods . During the course of expression of some of the PspA constructs, a shorter fragment was coexpressed due to translational pausing and subsequent secondary translation initiation . Two of the constructs, PspA-206 and PspA-272, were also crystallized allowing for the initiation of a structural elucidation of PspA .

Colloids Surf B Biointerfaces, 2001 Feb 1, 20(2), 189 - 194
Lack of effect of an externally applied electric field on bacterial adhesion to glass; Poortinga AT et al.; Deposition to glass of Streptococcus salivarius HB-C12 and Staphylococcus epidermidis 3399 in a parallel plate flow chamber in the absence and presence of an externally applied electric field has been studied experimentally . No effect on bacterial adhesion, including initial deposition rates, numbers of adhering bacteria after 4 h, spatial distributions of adhering bacteria and air bubble induced detachment, was found . A theoretical analysis shows that electric fields applied over a 150 microm thin glass substratum do not have a sufficiently strong effect on its surface potential to influence bacterial adhesion.

Colloids Surf B Biointerfaces, 2001 Feb 1, 20(2), 105 - 117
Electrostatic interactions in the adhesion of an ion-penetrable and ion-impenetrable bacterial strain to glass; Poortinga AT et al.; Deposition to glass of Streptococcus salivarius HB-C12 and Staphylococcus epidermidis 3399 in a parallel plate flow chamber has been studied as a function of ionic strength . Electrophoretic mobility measurements revealed that S . epidermidis 3399 possesses a thick ion-penetrable layer, probably associated with its encapsulation, while S . salivarius HB-C12 has an ion-impenetrable surface . Streaming potential measurements indicated that also the glass surface was covered with a relatively thin, ion-penetrable layer . Theoretical initial deposition rates of both strains to glass were obtained by numerically solving the convective-diffusion equation, while accounting for the ion-penetrability of the interacting surfaces . Experimentally, the initial deposition rate of the ion-penetrable strain S . epidermidis 3399 was found to be higher and less dependent on ionic strength than of the ion-impenetrable S . salivarius HB-C12, in accordance with theoretical expectations . Agreement between theoretical and experimental deposition rates could be obtained when glass was considered ion-penetrable when interacting with the ion-penetrable organism S . epidermidis 3399, while glass behaved as an ion-impenetrable surface when interacting with the ion-impenetrable S . salivarius HB-C12 . Probably, interaction with an ion-impenetrable strain drives the diffuse double layer charges into the limited volume of the thin ion-penetrable layer on the glass, readily filling it up and making it appear ion-impenetrable . During interaction of glass with another ion-penetrable surface, as of S . epidermidis 3399, diffuse double layer charges move into both ion-penetrable surfaces, resulting in a much lower mobile charge density in the ion-penetrable layer on the glass which consequently continues to behave as ion-penetrable.

J Infect Dis, 2001 Jan 1, 183(1), 70 - 7 Epub 2000 Nov 15.
Efficacy of recombinant human granulocyte colony-stimulating factor in a murine model of pneumococcal pneumonia: effects of lung inflammation and timing of treatment; Dallaire F et al.; The effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) in a murine model of pneumococcal pneumonia was examined . Intranasal inoculations were 10(7) cfu/mouse (high inoculum) and 5 x 10(4) cfu/mouse (low inoculum) of Streptococcus pneumoniae, which induced severe or mild lung inflammation, respectively . With the low inoculum, rhG-CSF significantly improved survival when initiated 24 h or 10 min before, but not when initiated 24 h after, infection . Pretreatment with rhG-CSF significantly increased myeloperoxidase (MPO) activity in lungs 8 h after the infection and increased circulating neutrophil count 24, 48, and 72 h after infection . In contrast, rhG-CSF did not improve survival of animals infected with the high inoculum and did not increase MPO activity or neutrophil count in blood over those of sham-treated controls . These data strongly suggest that the severe inflammatory response typically observed in pneumococcal pneumonia recruits a maximum number of neutrophils in the lungs and thus masks the beneficial effect of rhG-CSF.

Aesthetic Plast Surg, 2000 Sep-Oct, 24(5), 344 - 7
Rapidly fatal necrotizing fasciitis after aesthetic liposuction; Heitmann C et al.; Necrotizing fasciitis (NF) is a rapidly progressive soft tissue infection involving primarily the superficial fascia and subcutaneous tissue . The disease is caused by Streptococcus pyogenes or synergistic infection of anaerobic and facultative anaerobic bacteria . Further characteristics are severe, intolerable pain and a mortality rate of 30 to 50% . The NF can be initiated after surgical procedures, minor trauma, trivial scratches, in the setting of a chronic wound, or even in apparently intact skin . The age of the patient is not relevant for the prognosis of NF . As it is shown in this reported case, a young and previously healthy patient died after aesthetic liposuction in the course of a NF . Necrotizing fasciitis is a rare disease, therefore, it is important to review its diagnostic and clinical features, because only early diagnosis and prompt, radical surgery improves the survival rate.

Thorax, 2000 Dec, 55(12), 1040 - 5
A national confidential enquiry into community acquired pneumonia deaths in young adults in England and Wales . British Thoracic Society Research Committee and Public Health Laboratory Service; Simpson JC et al.; BACKGROUND: The aim of this study was to describe the frequency, causal pathogens, management, and outcome of a population of young adults who died from community acquired pneumonia (CAP) . METHODS: Pneumonia deaths in England and Wales in adults aged 15-44 were identified between September 1995 and August 1996 . Patients with underlying chronic illness including HIV infection were excluded . Clinical details for each case were collected from the hospital and general practitioner records . RESULTS: Death from CAP was identified in 27 previously well young adults (1.2 per million population per year) . Twenty were known to have consulted a GP for this illness . Nine received antibiotics before hospital admission . A causative pathogen was identified in 17 cases (Streptococcus pneumoniae in eight) . Bacteraemia was present in seven . All patients who reached a hospital ward received antibiotics (69% within two hours of admission) . The British Thoracic Society antibiotic guidelines for severe CAP were followed in only 10 cases . Cardiac arrest at home or on arrival at hospital occurred in six cases, one of whom was successfully resuscitated . Of the remaining 21 patients, 71% had two or more markers of severe CAP . All 22 who were admitted reached an intensive care unit, but 11 of these required transfer to another hospital for some aspect of intensive care . One third of patients died within 24 hours of presenting to the hospital . CONCLUSIONS: Death from CAP in previously fit young adults still occurs . While some deaths might be preventable by better patient management, most are unlikely to be preventable by current management practices.

Infect Immun, 2000 Dec, 68(12), 7141 - 3
Antigenicity, expression, and molecular characterization of surface-located pullulanase of Streptococcus pneumoniae; Bongaerts RJ et al.; A putative pullulanase-encoding gene from Streptococcus pneumoniae was identified by screening a genomic expression library with human convalescent-phase serum . The 3,864-bp gene encoded a 143-kDa protein . Surface location and pullulanase activity of the protein, designated SpuA, was demonstrated . SpuA was present in all investigated pneumococcal isolates of different serotypes . The spuA 5' end was highly conserved among clinical isolates except for a 75-bp region . The properties of SpuA reported here indicate that this novel immunogenic surface protein might have potential as a vaccine target.

Infect Immun, 2000 Dec, 68(12), 6807 - 18
Identification and immunogenicity of group A Streptococcus culture supernatant proteins; Lei B et al.; Extracellular proteins made by group A Streptococcus (GAS) play critical roles in the pathogenesis of human infections caused by this bacterium . Although many extracellular GAS proteins have been identified and characterized, there has been no systematic analysis of culture supernatant proteins . Proteins present in the culture supernatant of strains of serotype M1 (MGAS 5005) and M3 (MGAS 315) mutants lacking production of the major extracellular cysteine protease were separated by two-dimensional gel electrophoresis and identified by amino-terminal amino acid sequencing and interrogation of available databases, including a serotype M1 genome sequence . In the aggregate, amino-terminal amino acid sequence data for 66 protein spots were generated, 53 unique sequences were obtained, and 44 distinct proteins were identified . Sixteen of the 44 proteins had apparent secretion signal sequences and 27 proteins did not . Eight of the 16 proteins with apparent secretion signal sequences have not been previously described for GAS . Antibodies against most of the apparently secreted proteins were present in sera from mice infected subcutaneously with MGAS 5005 or MGAS 315 . Humans with documented GAS infections (pharyngitis, acute rheumatic fever, and severe invasive disease) also had serum antibodies reacting with many of the apparently secreted proteins, indicating that they were synthesized in the course of GAS-human interaction . The genes encoding four of the eight previously undescribed and apparently secreted culture supernatant proteins were cloned, and the proteins were overexpressed in Escherichia coli . Western blot analysis with these recombinant proteins and sera from GAS-infected mice and humans confirmed the immunogenicity of these proteins . Taken together, the data provide new information about the molecular aspects of GAS-host interactions.

Infect Immun, 2000 Dec, 68(12), 6542 - 53
Identification and characterization of the scl gene encoding a group A Streptococcus extracellular protein virulence factor with similarity to human collagen; Lukomski S et al.; Group A Streptococcus (GAS) expresses cell surface proteins that mediate important biological functions such as resistance to phagocytosis, adherence to plasma and extracellular matrix proteins, and degradation of host proteins . An open reading frame encoding a protein of 348 amino acid residues was identified by analysis of the genome sequence available for a serotype M1 strain . The protein has an LPATGE sequence located near the carboxy terminus that matches the consensus sequence (LPXTGX) present in many gram-positive cell wall-anchored molecules . Importantly, the central region of this protein contains 50 contiguous Gly-X-X triplet amino acid motifs characteristic of the structure of human collagen . The structural gene (designated scl for streptococcal collagen-like) was present in all 50 GAS isolates tested, which together express 21 different M protein types and represent the breadth of genomic diversity in the species . DNA sequence analysis of the gene in these 50 isolates found that the number of contiguous Gly-X-X motifs ranged from 14 in serotype M6 isolates to 62 in a serotype M41 organism . M1 and M18 organisms had the identical allele, which indicates very recent horizontal gene transfer . The gene was transcribed abundantly in the logarithmic but not stationary phase of growth, a result consistent with the occurrence of a DNA sequence with substantial homology with a consensus Mga binding site immediately upstream of the scl open reading frame . Two isogenic mutant M1 strains created by nonpolar mutagenesis of the scl structural gene were not attenuated for mouse virulence as assessed by intraperitoneal inoculation . In contrast, the isogenic mutant derivative made from the M1 strain representative of the subclone most frequently causing human infections was significantly less virulent when inoculated subcutaneously into mice . In addition, both isogenic mutant strains had significantly reduced adherence to human A549 epithelial cells grown in culture . These studies identify a new extracellular GAS virulence factor that is widely distributed in the species and participates in adherence to host cells and soft tissue pathology.

Antimicrob Agents Chemother, 2000 Dec, 44(12), 3447 - 50
Comparative In vitro activities of daptomycin and vancomycin against resistant gram-positive pathogens; Snydman DR et al.; The in vitro activity of daptomycin against 224 current gram-positive clinical isolates including vancomycin-resistant Enterococcus faecium (VREF), methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus spp . (MRSS), and penicillin-resistant Streptococcus pneumoniae (PRSP) was evaluated . The MICs at which 90% of isolates are inhibited for daptomycin and vancomycin, respectively, were as follows: MRSA, 1 and 2 microg/ml; MRSS, 1 and 4 microg/ml; PRSP, 1 and 0.5 microg/ml; and VREF, 2 and >64 microg/ml . Daptomycin was bactericidal against 82% of 17 VREF isolates . The antibacterial activity of daptomycin was strongly dependent on the calcium concentration of the medium . Daptomycin was active against all gram-positive cocci tested.

Antimicrob Agents Chemother, 2000 Dec, 44(12), 3395 - 401
Two new mechanisms of macrolide resistance in clinical strains of Streptococcus pneumoniae from Eastern Europe and North America; Tait-Kamradt A et al.; Resistance to macrolides in pneumococci is generally mediated by methylation of 23S rRNA via erm(B) methylase which can confer a macrolide (M)-, lincosamide (L)-, and streptogramin B (S(B))-resistant (MLS(B)) phenotype or by drug efflux via mef(A) which confers resistance to 14- and 15-membered macrolides only . We studied 20 strains with unusual ML or MS(B) phenotypes which did not harbor erm(B) or mef(A) . The strains had been isolated from patients in Eastern Europe and North America from 1992 to 1998 . These isolates were found to contain mutations in genes for either 23S rRNA or ribosomal proteins . Three strains from the United States with an ML phenotype, each representing a different clone, were characterized as having an A2059G (Escherichia coli numbering) change in three of the four 23S rRNA alleles . Susceptibility to macrolides and lincosamides decreased as the number of alleles in isogenic strains containing A2059G increased . Sixteen MS(B) strains from Eastern Europe were found to contain a 3-amino-acid substitution ((69)GTG(71) to TPS) in a highly conserved region of the ribosomal protein L4 ((63)KPWRQKGTGRAR(74)) . These strains formed several distinct clonal types . The single MS(B) strain from Canada contained a 6-amino-acid L4 insertion ((69)GTGREKGTGRAR), which impacted growth rate and also conferred a 500-fold increase in MIC on the ketolide telithromycin . These macrolide resistance mechanisms from clinical isolates are similar to those recently described for laboratory-derived mutants.

Eur J Biochem, 2000 Dec, 267(23), 6810 - 6
Covalent structure of mutacin 1140 and a novel method for the rapid identification of lantibiotics; Smith L et al.; The primary structure of the Streptococcus mutans lantibiotic mutacin 1140 was elucidated by NMR spectroscopy, mass spectrometry, and chemical sequencing . The structure is in agreement with other closely related lantibiotics, such as epidermin . A novel method was developed in which mutacin 1140 was chemically modified with sodium borohydride followed by ethanethiol, allowing the differentiation of the thioether-containing residues from the dehydrated residues . This double-labeling strategy provides a simple method to reliably identify all modified lantibiotic residues with a minimal amount of material . While NMR spectroscopy is still required to obtain thioether bridging patterns and thus the complete covalent structure, the double-labeling technique, along with mass spectrometry, provides most of the information in a fraction of the time required for a complete NMR analysis . Thus, with these new techniques lantibiotics can be rapidly characterized.

Pharmacotherapy, 2000 Nov, 20(11), 1310 - 7
Comparative in vitro activity and pharmacodynamics of five fluoroquinolones against clinical isolates of Streptococcus pneumoniae; Kays MB et al.; STUDY OBJECTIVE: To compare in vitro activity and pharmacodynamics of five fluoroquinolones against clinical isolates of Streptococcus pneumoniae . DESIGN: In vitro analysis . SETTING: University research laboratory . INTERVENTION: Minimum inhibitory concentrations (MICs) were determined for penicillin and five fluoroquinolones by E test for 201 S . pneumoniae isolates . Serum concentration-time profiles were simulated for the following regimens: ciprofloxacin 750 mg orally every 12 hours and 400 mg intravenously every 8 hours; levofloxacin 500 mg orally and intravenously every 24 hours; trovafloxacin 200 mg orally and intravenously every 24 hours; gatifloxacin 400 mg orally and intravenously every 24 hours; and clinafloxacin 200 mg orally and intravenously every 12 hours . MEASUREMENTS AND MAIN RESULTS: Free 24-hour areas under the serum concentration-time curves (AUC0-24) were calculated using the trapezoidal rule, and the average AUC0-24:MIC ratio was calculated for each regimen . Differences in ratios among agents were determined by analysis of variance (Scheffe post hoc test, p < 0.05) . For intravenous dosing, the average AUC0-24:MIC for gatifloxacin, clinafloxacin, trovafloxacin, ciprofloxacin, and levofloxacin was 146, 142, 122, 71, and 61, respectively . For both oral and intravenous regimens, gatifloxacin and clinafloxacin ratios were significantly greater than those for trovafloxacin, levofloxacin, and ciprofloxacin (p < or = 0.007) . Ratios for trovafloxacin were significantly greater than those for levofloxacin and ciprofloxacin (p < 0.0001), and levofloxacin and ciprofloxacin ratios were not significantly different from each other . CONCLUSION: Gatifloxacin and clinafloxacin achieve significantly higher AUC0-24:MIC ratios for S . pneumoniae than trovafloxacin, levofloxacin, and ciprofloxacin . Large comparative studies are necessary to determine the clinical significance of these findings.

Int J Mol Med, 2000 Dec, 6(6), 699 - 703
Streptococcus anginosus in head and neck squamous cell carcinoma: implication in carcinogenesis; Tateda M et al.; It has been suggested that Helicobacter pylori (H . pylori) infection might be associated with not only gastric ulcers but also gastric malignancies . Recently, it was reported that the Streptococcus anginosus (S . anginosus) DNA sequence was found in DNA samples extracted from esophageal cancers . Because smoking and alcohol abuse are regarded as risk factors for both esophgeal cancer and head and neck cancer, infection of S . anginosus might be associated with carcinogenesis of head and neck cancer . To investgate the involvement of S . anginosus infection in head and neck cancer, we analyzed 217 DNA samples prepared from head and neck squamous cell carcinomas . We performed PCR analysis with S . anginosus-16S ribosomal DNA-specific primers, and Southern blot analysis . For detection of S . anginosus in the oral and pharyngeal cavities, we used oropharyngeal bacteriological culture and PCR analysis of gingival smears of the patients . By PCR analysis, the S . anginosus DNA sequence was found in 217 out of 217 (100%) DNA samples obtained from head and neck cancers . By Southern blot analysis, positive bands were detected in 41 out of 125 (33%) samples . We could find no S . anginosus colony in oropharyngeal bacteriological culture dishes of 53 patients with and without head and neck cancer . On the other hand, we found the S . anginosus DNA fragment in 8 out of 8 DNA samples obtained from gingival smears by PCR analysis . These data indicate that the upper aerodigestive environment of the patients permitting S . anginosus infection was implicated in the carcinogenesis of head and neck squamous cell carcinoma.

Cell Mol Biol (Noisy-le-grand), 2000 Nov, 46(7), 1297 - 303
Red light kills bacteria via photodynamic action; Konig K et al.; With the increase in the number of antibiotic resistant strains of microorganism, the search for alternative treatments of microbial infections becomes all the more important . We report a novel method for bacterial inactivation based on the optical excitation of the naturally occurring (endogenous) photosensitzing porphyrins by red light . In particular, the pathogenic Gram-positive porphyrin producing ATCC strains Propionibacterium acnes, Actinomyces odontolyticus and Porphyromonas gingivalis were investigated . Sensitive autofluorescence spectroscopy revealed that these bacteria naturally synthezise the fluorescent photosensitizer protoporphyrin IX . In addition, bacterial plaque samples of periodontitis patients were studied . Non-labeled fluorescent bacterial colonies were exposed to red light at 632.8 nm, 100 mW/cm2 light intensity and 360 J/cm2 energy density using a helium-neon laser . The survival rate after a single phototreatment with red light was found to be 0.58 +/- 0.09 in the case of Propionibacterium acnes, 0.30 +/- 0.04 in Actinomyces odontolyticus and 0.59 +/- 0.10 in Porphyrormonas gingivalis compared to non-exposed bacteria suspensions . No photoeffect was found for the bacterium Streptococcus mutans which exhibited no detectable porphyrin autofluorescence . Red-light exposed plaque samples of patients showed significant reduction of colony forming units by 50% as well as a pronounced photoeffect on the pigmented species Prevotella intermedia . Taken together, these results suggest the treatment with red light can be potentially employed as an therapeutic method to inactivate certain pathogenic strains of porphyrin producing bacteria without the use of external photosensitizers.

Mayo Clin Proc, 2000 Nov, 75(11), 1161 - 8
Antimicrobial resistance in Streptococcus pneumoniae: implications for patients with community-acquired pneumonia; Chenoweth CE et al.; Streptococcus pneumoniae is the leading cause of community-acquired pneumonia . During the past decade, the prevalence of penicillin resistance in S pneumoniae has increased dramatically, with resistance rates approaching 45% in some areas of the United States . Streptococcus pneumoniae has also acquired resistance to other commonly used antimicrobials, including cephalosporins, macrolides, and trimethoprim-sulfamethoxazole . While vancomycin and the newer quinolones are currently highly active against most strains of S pneumoniae, reduced susceptibilities to these agents have been identified in some strains . Prior use of antimicrobial agents is the major risk factor for colonization and infection with antibiotic-resistant strains . beta-Lactam antibiotics remain the treatment of choice for infections caused by susceptible S pneumoniae . The optimum therapy for penicillin-resistant strains remains unclear . Appropriate empirical therapy for patients with community-acquired pneumonia depends in part on the community-specific resistance patterns of S pneumoniae to various antibiotics . In this article, we provide an overview of the development of S pneumoniae resistance to commonly used antibiotics and discuss the implications of the development of resistance on treatment decisions.

J Vet Med B Infect Dis Vet Public Health, 2000 Oct, 47(8), 581 - 9
Milk leucocyte population patterns in bovine udder infection of different aetiology; Leitner G et al.; This study compared the different leucocyte populations in milk from udders infected with different mastitic pathogens and in different stages of infection . Milk samples were collected from quarters free of intramammary infection, acutely infected with Escherichia coli or Staphylococcus aureus and chronically infected with S . aureus, coagulase-negative staphylococci (CNS) or Streptococcus dysgalactiae . Udder bacteriological status was confirmed after three consecutive bacteriological examinations from weekly quarter milk samples . At the time of the trial, milk samples were tested for somatic cell count (SCC) and differential cell count by both light microscopy (LM) and flow cytometry . Monoclonal antibody (mAb) CD11a/CD18 was used in order to differentiate between leucocytes and epithelial cells when tested by flow cytometry . Udder quarters free of intramammary infection had a mean SCC lower than 107 x 10(3) cells/ml in which the epithelial cells were the main cell type followed by polymorphonuclear cells (PMNs), while macrophages and lymphocytes had a lower concentration . Only 56% of the cells were labelled with the mAb anti-CD11a/CD18 . In either acute E . coli- or S . aureus-infected quarters, SCC were significantly higher (P < 0.0001) than in samples from the time of inoculation, with over 90% of the cells labelled with the mAb anti-CD11a/CD18 . The main cell type was neutrophils . In chronically infected cows, differences in SCC and in leucocyte patterns were found between infecting pathogens as well as between quarters harbouring the same pathogen . In all the chronically infected quarters, SCC was significantly higher (P < 0.05) than in uninfected ones . The distribution of the leucocyte patterns in the quarters infected with S . dysgalactiae did not differ from that in quarters with acute infection with both E . coli and S . aureus . In the cows chronically infected with S . aureus or CNS, the proportion of PMN was higher but not significantly different from quarters free of intramammary infection, while epithelial cells were significantly lower (P < 0.05) . The T lymphocytes bearing CD4+ or CD8+ were significantly higher in quarters chronically infected with S . aureus than in quarters free of intramammary infection and in quarters acutely infected with either E . coli or S . aureus . In all samples B cells were negligible.

Rev Med Interne, 2000 Oct, 21(10), 844 - 53
{Details of meningitis in the elderly}; Le Moal G et al.; PURPOSE: Elderly patients being more at risk for infections than younger people, this study was aimed at defining the epidemiological and clinical features of meningitis in this population, with the objective of improving diagnosis and management . METHODS: Over a period of 10 years, all cases admitted to an infectious diseases unit for acute meningitis were reviewed . Patients infected by human immunodeficiency virus (HIV), fungi meningitis, and who were younger than 15 years of age were excluded from the study . According to age, two groups were constituted (group A: < 65 years and group B: > or = 65 years) and compared . RESULTS: One hundred fifty-two patients were included in the study: 110 (72.4%) in group A (29 with bacterial and 81 with viral meningitis) and 42 (26.7%) in group B (32 with bacterial and ten with viral meningitis); the mean age was, respectively, 32.7 +/- 12.9 years (range: 15-61 years) and 75.9 +/- 7.6 years (range: 65-94 years) . Diagnosis was less frequently evoked in the elderly (n = 11; 26%) than in younger patients (n = 78; 71%) (P < 0.001) . Streptococcus pneumoniae, Listeria monocytogenes, and herpes simplex virus were the three most common causal agents in group B . Confusion was the most common symptom among the elderly (88 vs . 17%; P < 0.001) . The mortality rate was more important in group B than in group A (11.9 vs . 2.7%; P = 0.04) . CONCLUSIONS: Diagnosis of acute meningitis is difficult and must be evoked in the presence of any new neurologic sign . Cranial computerized tomography should not delay lumbar puncture, except in the presence of focal neurologic symptoms . Antimicrobial therapy takes into account the bacterial epidemiology.

J Biol Chem, 2001 Feb 23, 276(8), 5714 - 9 Epub 2000 Nov 13.
Structural analysis of the protein/lipid complexes associated with pore formation by the bacterial toxin pneumolysin; Bonev BB et al.; Pneumolysin, a major virulence factor of the human pathogen Streptococcus pneumoniae, is a soluble protein that disrupts cholesterol-containing membranes of cells by forming ring-shaped oligomers . Magic angle spinning and wideline static (31)P NMR have been used in combination with freeze-fracture electron microscopy to investigate the effect of pneumolysin on fully hydrated model membranes containing cholesterol and phosphatidylcholine and dicetyl phosphate (10:10:1 molar ratio) . NMR spectra show that the interaction of pneumolysin with cholesterol-containing liposomes results in the formation of a nonbilayer phospholipid phase and vesicle aggregation . The amount of the nonbilayer phase increases with increasing protein concentration . Freeze-fracture electron microscopy indicates the coexistence of aggregated vesicles and free ring-shaped structures in the presence of pneumolysin . On the basis of their size and analysis of the NMR spectra it is concluded that the rings are pneumolysin oligomers (containing 30-50 monomers) complexed with lipid (each with 840-1400 lipids) . The lifetime of the phospholipid in either bilayer-associated complexes or free pneumolysin-lipid complexes is > 15 ms . It is further concluded that the effect of pneumolysin on lipid membranes is a complex combination of pore formation within the bilayer, extraction of lipid into free oligomeric complexes, aggregation and fusion of liposomes, and the destabilization of membranes leading to formation of small vesicles.

Forensic Sci Int, 2001 Jan 15, 115(3), 199 - 205
Post-traumatic meningitis: histomorphological findings, postmortem microbiology and forensic implications; Matschke J et al.; Infections of the leptomeninges with the infectious agent gaining access to the intracranial compartment by traumatic means are termed post-traumatic . In cases with fatal outcome, the manner of death has to be classified as non-natural . Six cases of post-traumatic meningitis as the cause of death from the archives of the Institute of Legal Medicine in Hamburg, Germany with histological and microbiological investigations are presented . There were all males, age varying between 24 and 90 years (mean 58 years); range of the interval between original trauma and beginning of symptoms was 2 days up to 8 years; in 50% of the cases meningeal swabs yielded Streptococcus pneumoniae . Findings concerning origin and mechanism of post-traumatic meningitis as well as microbiological studies are compared with selected cases from the literature.

Int J Pediatr Otorhinolaryngol, 2000 Nov 30, 56(1), 33 - 40
Acute mastoiditis in children: review of the current status; Spratley J et al.; BACKGROUND: acute mastoiditis is the most common intratemporal complication of otitis media . Its management is still a challenge due to potentially serious consequences . This study was designed to evaluate the recent experience with pediatric acute mastoiditis at our institution and to determine if the incidence of this entity is changing over time . MATERIAL AND METHODS: retrospective review of records of children with acute mastoiditis treated at the hospital of the Medical School at the University of Porto, Portugal, between July 1993 and June 1998 . Criteria for the diagnosis of acute mastoiditis were postauricular swelling and erythema, protrusion of the auricle, and evidence of co-existent or recent otitis media . RESULTS: 43 patients fulfilled the entry criteria . Most were boys (69%) . Ages ranged from 8 months to 14 years and 4 months; infants represented 40% of the total . Acute mastoiditis was the first recognized sign of otitis media in 48% of patients . More recent years of the study saw an increase in the number of children referred with acute mastoiditis . Upon admission, 56% were under antibiotic treatment, with an average intake of 5.8 days . All patients were hospitalized; 26 cases recovered after intravenous antibiotics plus myringotomy, and the rest required an additional surgical procedure . The most common organisms recovered from cultures were Streptococcus pneumoniae and Streptococcus pyogenes . In our series, associated complications occurred in 13.9%; facial paralysis in one, and involvement of the central nervous system in five . CONCLUSIONS: pediatric acute mastoiditis continues to be a potentially dangerous infection in the antibiotic era . The incidence of this complication may be increasing recently in the community studied . Great care is required of clinicians to reach an early diagnosis in order to promote adequate management and prevent inherently severe complications.

J Bacteriol, 2000 Dec, 182(23), 6798 - 805
Distribution of the mosaic structured murM genes among natural populations of Streptococcus pneumoniae; Filipe SR et al.; The presence and sequence variation of the murM gene were studied in a large collection (814 strains) of genetically diverse Streptococcus pneumoniae isolates, which included 27 different serogroups and both penicillin-resistant (423 isolates, 67 pulsed-field gel electrophoretic {PFGE} types) and intermediately penicillin-resistant (165 isolates, 66 PFGE types) and penicillin-susceptible (226 isolates, 135 PFGE types) strains . Diversity of the murM sequences was tested by hybridization with mainly two kinds of probes: one derived from the amplification of the nucleotide sequence between nucleotides 201 and 624 in the penicillin-susceptible laboratory strain R36A (murMA probe) and a second probe that amplified the comparable, highly divergent sequence in the penicillin-resistant strain Pen6 (murMB probe) . The great majority of the strains (761 of 814), including both penicillin-susceptible and penicillin-resistant isolates, reacted exclusively with the murMA probe . A smaller group of penicillin-resistant strains (48 of 814 isolates) reacted only with the murMB DNA probe, and an additional 5 isolates reacted with both probes . High-pressure liquid chromatography analysis of the peptidoglycan of strains hybridizing with murMB showed that they invariably contained an increased proportion of branched peptides . Complete sequencing of murM from a group of penicillin-resistant isolates allowed the identification of a number of different murMB alleles that differed in the length and exact position of the divergent (Pen6 type) sequences within the particular murM . The close similarity of these divergent sequences in the various murM alleles suggests a possible common heterologous origin.

Clin Infect Dis, 2000 Nov, 31(5), E23 - 5
Group B streptococcal meningitis complicating elective abortion: report of 2 cases; Deziel PJ et al.; The incidence of invasive disease due to Streptococcus agalactiae (group B streptococcus {GBS}) in adults is on the rise; however, meningitis in adults due to GBS remains rare . We report 2 cases of GBS meningitis complicating elective abortion, 1 of which was a septic incomplete abortion . Only 1 case of bacterial meningitis complicating elective abortion has been reported previously.

J Vet Med Sci, 2000 Oct, 62(10), 1053 - 7
A 10-year survey of antimicrobial susceptibility of streptococcus suis isolates from swine in Japan; Kataoka Y et al.; A number of 689 Streptococcus suis isolates collected nationwide from diseased and healthy pigs from 1987 to 1996 were surveyed for antibiotic susceptibilities to 11 drugs . No isolates resistant to amoxicillin, chloramphenicol, and sulfamethoxazole/trimethoprim were found . Isolates were highly susceptible to penicillins (penicillin G, ampicillin, and amoxicillin) except cloxacillin . They were not susceptible to tetracycline, streptomycin, and kanamaycin (MIC90 50 microg/ml, > or = 100 microg/ml, and > or = 100 microg/ml, respectively) . Multiple-resistant isolates (> or = 3 antimicrobial agents) were found in 20.3% of all isolates tested.

Presse Med, 2000 Oct 7, 29(29), 1601 - 2
{Streptococcus intermedius Lemierre syndrome: an unusual association}; Chemlal K et al.; BACKGROUND: Lemierre's syndrome is a potentially severe though uncommon entity consisting in septic emboli from an internal jugular vein thrombus after oropharyngeal infection . CASE REPORT: A 21-year-old man initially treated for acute pharnygeal infection developed fever and lower chest pain related to multiple pulmonary abscesses . The diagnosis of Lemierre's syndrome was retained due to the association of extensive thrombus formation in the internal jugular vein and Streptococcus intermedius septicemia . DISCUSSION: Lemierre's syndrome is a classical entity whose frequency is probably underestimated . The causal agent is not always an anaerobic germ . Use of anticoagulants is controversial.

Semin Arthritis Rheum, 2000 Oct, 30(2), 121 - 6
Pyarthrosis in patients with rheumatoid arthritis: a detailed analysis of 10 cases and literature review; Nolla JM et al.; OBJECTIVES: 1) To analyze the clinical features and outcome of patients with rheumatoid arthritis and pyarthrosis seen in a rheumatology department during a 9-year period; 2) To review the available literature about this association in the last decade . METHODS: From the database of our department, we collected all hospitalized cases of infectious arthritis in native joints between January 1990 and December 1998 . In 10 cases (27%), pyarthrosis occurred in patients with rheumatoid arthritis . A detailed analysis of each patient was performed . The literature was reviewed by using MEDLINE from 1990 to 1999 . RESULTS: The mean age of patients was 63.2 years; six were female . Most patients had long-standing disease and poor functional class, and all received glucocorticoid treatment . Mean diagnostic delay was 7.3 days . Causative organisms were Staphylococcus aureus (4 cases), gram-negative bacilli (3 cases), anaerobic bacteria (2 cases), and Streptococcus pneumoniae (n = 1) . Two patients died . In all but two patients who survived, joint function worsened . CONCLUSIONS: Rheumatoid arthritis is a relevant host-related risk factor for septic arthritis . Pyarthrosis in these patients is associated with considerable morbidity and mortality.

J Infect Dis, 2000 Dec, 182(6), 1702 - 11 Epub 2000 Oct 18.
Characterization of a mouse-passaged, highly encapsulated variant of group A streptococcus in in vitro and in vivo studies; Ravins M et al.; JRS4(HE), a highly encapsulated, mouse-passaged variant of group A streptococcal strain JRS4, was characterized . The mucoid phenotype of JRS4(HE) was preserved after extensive passage in vitro . The level and size of csrRS transcript in JRS4(HE) was similar to that of JRS4, yet JRS4(HE) expressed high levels of has and sagA and exhibited an increased activity of streptolysin S . These findings indicate that the CsrRS repressor system was inactive in JRS4(HE) . JRS4(HE) adhered to HEp-2 cells at the stationary phase but did not internalize these cells . At midlogarithmic phase, JRS4(HE) neither adhered to nor internalized cells, because of an increased amount of hyaluronic acid . Mice injected subcutaneously with JRS4(HE) developed large, deep necrotic lesions . In contrast, mice challenged with JRS4 developed small, superficial lesions . Despite the use of a high inoculum, mice challenged with JRS4(HE) did not develop a lethal bacteremic infection . It is concluded that inactivation of CsrRS in vivo is insufficient to cause a spreading necrotic disease.

J Infect Dis, 2000 Dec, 182(6), 1694 - 701 Epub 2000 Nov 08.
Immunization of humans with recombinant pneumococcal surface protein A (rPspA) elicits antibodies that passively protect mice from fatal infection with Streptococcus pneumoniae bearing heterologous PspA; Briles DE et al.; Pneumococcal surface protein A (PspA), a cross-reactive protein expressed by all pneumococci, is known to elicit an antibody in animals that can passively protect mice from infection with Streptococcus pneumoniae . A phase I trial with recombinant PspA showed the protein to be immunogenic in humans . Pre- and postimmune serum samples from this trial were examined, and human antibody to PspA could protect mice from pneumococcal infection . The serum samples of subjects immunized twice with 125 microg of PspA had >100 times as much antibody per milliliter as was required to consistently protect mice from fatal infection (1.3 microg/dose) . At least 98% of PspAs fall into PspA sequence/serologic families 1 or 2 . Human antibodies elicited by a family 1 PspA protected against infection with S . pneumoniae expressing either family 1 or 2 PspAs and with strains of all 3 capsular types tested: 3, 6A, and 6B . These studies suggest that PspA may have efficacy as a human vaccine.

Arch Biochem Biophys, 2000 Oct 15, 382(2), 303 - 9
Purification and characterization of the RecA protein from Streptococcus pneumoniae; Steffen SE et al.; Streptococcus pneumoniae is a naturally transformable bacterium that is able to take up single-stranded DNA from its environment and incorporate the exogenous DNA into its genome . This process, known as transformational recombination, is dependent upon the presence of the recA gene, which encodes an ATP-dependent DNA recombinase whose sequence is 60% identical to that of the RecA protein from Escherichia coli . We have developed an overexpression system for the S . pneumoniae RecA protein and have purified the protein to greater than 99% homogeneity . The S . pneumoniae RecA protein has ssDNA-dependent NTP hydrolysis and NTP-dependent DNA strand exchange activities that are generally similar to those of the E . coli RecA protein . In addition to its role as a DNA recombinase, the E . coli RecA protein also acts as a coprotease, which facilitates the cleavage and inactivation of the E . coli LexA repressor during the SOS response to DNA damage . Interestingly, the S . pneumoniae RecA protein is also able to promote the cleavage of the E . coli LexA protein, even though a protein analogous to the LexA protein does not appear to be present in S . pneumoniae.

Kansenshogaku Zasshi, 2000 Sep, 74(9), 703 - 8
{Retrospective studies of penicillin-resistant pneumococcal acute otitis media in infants and children--the treatment of tympanostomy tube insertion}; Uno Y; Retrospective studies of treatment of tympanostomy tube insertion in infantile otitis prone patients caused by Penicillin-resistant Streptococcus pneumoniae were performed . Insertion of the Koken B type tympanostomy tube was performed with local anesthesia . The results were as follows: 1 . In 25 otitis prone children, 19 (76.0%) were not recognized as having otitis media and otorrhea, but 6 (24.0%) were recognized . 2 . Of the 25 children, 20 had the tympanostomy tube removed . Eighteen were not recognized as having otitis media and otorrhea during tympanostomy tube insertion, but 2 were recognized . 3 . In 18 children, 2 developed otitis media after tympanostomy tube removal, but the other 16 children did not . 4 . In 2 children who were recognized as having otitis media and otorrhea during tympanostomy tube insertion, 1 child actually had otitis media, whereas the other did not . 5 . From this study, tympanostomy tube insertion is a useful method for treating the otitis prone children caused by Penicillin-resistant Streptococcus pneumoniae.

Anal Biochem, 2000 Nov 15, 286(2), 238 - 46
Open sandwich enzyme-linked immunosorbent assay for the quantitation of small haptens; Suzuki C et al.; The quantitation of low-molecular-weight haptens has been difficult with conventional sandwich immunoassays due to their small size . Many researchers have attempted to develop sandwich assays for haptens due to the significant advantages of the sandwich format over competitive assays including greater dynamic range, ease of automation, and sensitivity . Here we apply the open-sandwich ELISA (OS-ELISA), an immunoassay based on antigen-dependent stabilization of antibody variable regions (V(H) and V(L) domains), to hapten quantitation . Two fusion proteins, the high-affinity mutant V(H) domain from anti-4-hydroxy-3-nitrophenacetyl (NP) antibody B1-8 tethered with Escherichia coli alkaline phosphatase (V(H)(W33L)-PhoA) and the V(L) domain from the same antibody tethered with Streptococcus sp . protein G, were made . These fusion proteins when added together achieved Fv reassociation consequent to the addition of NP . Signal was generated in a direct relationship to the NP concentration with better sensitivity compared with competitive immunoassay, demonstrating this assay to be a quick noncompetitive alternative to the conventional assays for small compounds, such as environmental pollutants, drugs of abuse, and therapeutic drugs . With our previous demonstration that the OS-ELISA works well with large proteins, the OS-ELISA becomes the first practical immunoassay approach capable of quantifying any molecule regardless of their size .

Microbiology, 2000 Nov, 146 ( Pt 11), 2785 - 92
Mitogenic factor (MF) is the major DNase of serotype M89 Streptococcus pyogenes; Sriskandan S et al.; To investigate the role of mitogenic factor (MF) in streptococcal pathogenesis, the structural gene (mf) encoding this protein was disrupted in a clinical isolate of Streptococcus pyogenes H293, to yield the isogenic mutant H363 . Growth in enriched broth and on blood agar was unaffected by disruption of mf . Cell-free broth supernatants from H293 and H363 demonstrated identical promitogenic activities when co-incubated with human peripheral blood mononuclear cells, even when diluted 100000-fold, showing that MF is not a major streptococcal mitogen compared with other secreted superantigens . Disruption of mf resulted in complete loss of DNase B production and detectable DNase activity in H363 compared with the parent strain, confirming that the single gene mf, which is present in all group A streptococcal M serotypes studied, encodes DNase B . Despite loss of DNase activity, the virulence of S . pyogenes in a mouse model of necrotizing fasciitis and myositis was unaffected.

Intern Med, 2000 Nov, 39(11), 991 - 3
A diabetic patient with scrotal subcutaneous abscess; Takemura T et al.; A 51-year-old type 2 diabetic patient with a scrotal subcutaneous abscess is reported . He was diagnosed as having diabetes mellitus five years earlier . He had left scrotal swelling and pain with granulocytosis, elevated C-reactive protein and hyperglycemia . He was successfully treated with incision and drainage (Streptococcus agalactiae was identified in the pus), debridement, antibiotics, immunoglobulin and insulin . This case resembled Fournier's gangrene, an infective necrotizing fasciitis of the perineal, genital or perianal regions . Diabetes mellitus is a basic disorder often associated with Fournier's gangrene . Scrotal subcutaneous abscess should be prevented from progressing to Fournier's gangrene with early and appropriate treatment.

Aust N Z J Obstet Gynaecol, 2000 Aug, 40(3), 338 - 40
Universal antenatal group B streptococcus screening? The opinions of obstetricians and neonatologists within Australia; McLaughlin K et al.; Group B streptococcus (GBS) is the leading infectious cause of morbidity and mortality in Australian newborns . Although intrapartum chemoprophylaxis is recommended to reduce the risk of neonatal GBS transmission and disease, controversy exists as to the best method to select women 'at risk' for this treatment . Our study aimed to survey the opinions of obstetricians and neonatologists currently in practice in Australia on GBS screening and treatment . Of the 488 obstetricians and 68 neonatologists currently in practice who responded to the survey, 271 obstetricians (56%) and 40 neonatologists (61%) supported universal antenatal screening . Of those respondents who did not support a universal antenatal screening policy, 196 (93%) and 24 (92%) of the obstetricians and neonatologists respectively, supported antenatal screening based on risk factors . This diversity in practitioner opinion highlights the lack of certainty in the literature as to the best management strategy to prevent neonatal GBS sepsis.

Ann Trop Paediatr, 2000 Sep, 20(3), 193 - 8
Bacterial infection in children with HIV: a prospective study from Cape Town, South Africa; Westwood AT et al.; Invasive bacterial infection in children infected with the human immunodeficiency virus (HIV) is common . South African data on this problem are limited . Over 1 year we prospectively studied 108 HIV-infected children hospitalized for 136 presumed infective episodes . Blood culture was positive in 24.8% of episodes . Streptococcus pneumoniae predominated (14/30 positive blood cultures); one-third of isolates showed resistance to penicillin . Acute lower respiratory tract infection accounted for 44% of clinical diagnoses, a bacterial cause being established for 23.8% of these . Age and stage of HIV infection did not influence the likelihood of a positive culture . A high proportion of presumed infective episodes requiring hospitalization of young HIV-infected children have a bacterial cause . Blood culture appears to be a useful method of obtaining the microbiological information required to focus antibiotic therapy.

FEMS Immunol Med Microbiol, 2000 Nov, 29(3), 183 - 5
Immune-protective antibodies against capsular polysaccharides do not affect natural competence of Streptococcus pneumoniae: implications for current conjugate vaccination strategies?
Overweg K, Sluijter M, Srodzinski M, de Groot R, Hermans PW.
We studied the effect of opsonization of Streptococcus pneumoniae with capsular antibodies on horizontal transfer of DNA . Opsonization did not inhibit DNA uptake . This suggests that horizontal transfer of capsular genes, which is an important escape mechanism of the pathogen, remains a potential threat for the efficacy of conjugate vaccination.

MMWR Morb Mortal Wkly Rep, 2000 Oct 20, 49(41), 936 - 40
Hospital-based policies for prevention perinatal Group B streptococcal disease--United States, 1999; Antibiotic resistance in Streptococcus pneumoniae; Canberra Clinical School, Canberra Hospital, ACT . peter.collignon@act.gov.au

Resistance to penicillin and other antibiotics in Streptococcus pneumoniae has emerged in Australia and around the world in the past decade, and appears to be worsening (e.g., rates of penicillin resistance in Australia rose from 1% in 1989 to 25% in 1997) . In Australia, the only oral antibiotic able to treat respiratory infections caused by some multiresistant strains is high-dose amoxycillin . If these strains increase in prevalence, then treatment failures for relatively minor infections (e.g., otitis media) are likely to become common, resulting in repeat antibiotic courses or hospitalisation for parenteral therapy . Therapy for meningitis caused by penicillin-sensitive pneumococcal strains remains high-dose benzylpenicillin, but empirical treatment while awaiting culture and sensitivity results is problematic; neither penicillin nor third-generation cephalosporins cover all strains . Therefore, many authorities recommend vancomycin, usually combined with a third-generation cephalosporin, for treating presumptive or proven pneumococcal meningitis pending penicillin-susceptibility results . As almost all readily available oral antibiotics in Australia select for resistant strains of pneumococci, multiresistant strains will increase in prevalence unless unnecessary antibiotic use and prescription volumes are reduced substantially in the next few years.

Med J Aust, 2000 Oct 2, 173 Suppl, S36 - 40
Are current recommendations for pneumococcal vaccination appropriate for Western Australia? The Vaccine Impact Surveillance Network--Invasive Pneumococcal Study Group; Invasive pneumococcal disease in the population of Victoria; Microbiological Diagnostic Unit, University of Melbourne, VIC . g.hogg@mdu.unimelb.edu.au

OBJECTIVES: To estimate morbidity and mortality rates for invasive Streptococcus pneumoniae (pneumococcal) disease in the non-Indigenous population of Victoria . DESIGN AND SETTING: Survey using data from a statewide voluntary laboratory surveillance scheme (1989-1998), statewide hospital discharge database (1995-1998), medical records of notified patients (1994-1995) and serotyping of notified isolates (1994-1998) . MAIN OUTCOME MEASURES: Incidence of pneumococcal bacteraemia and pneumonia; predisposing factors; serotypes of isolates . RESULTS: Minimum estimates of annual incidence of invasive disease, based on laboratory surveillance data for 1995-1998, were 59 per 100,000 for children aged < 2 years, 25 per 100,000 for people aged > or = 65 years, and 8 per 100,000 overall . Annual incidence of pneumococcal pneumonia, calculated from hospital discharge data, was 99 per 100,000 for those aged > or = 65 years . Manifestations of invasive pneumococcal disease varied with age, with meningitis more common in infants, and pneumonia most common in older patients . A predisposing factor for pneumococcal infection was present in 48% of patients . Most isolates from infants (83%) belonged to serotypes in the proposed seven-valent infant vaccine, and 91% of isolates from people aged > or = 2 years belonged to serotypes in the current 23-valent adult vaccine . CONCLUSIONS: S . pneumoniae continues to be a major cause of morbidity and mortality in young children and the elderly in Victoria . More widespread use of the currently available pneumococcal vaccine in adults and introduction of an effective vaccine for infants should greatly reduce incidence of the disease.

J Antimicrob Chemother, 2000 Nov, 46(5), 843 - 6
Safety and efficacy of recombinant granulocyte colony-stimulating factor as an adjunctive therapy for Streptococcus pneumoniae meningitis in non-neutropenic adult patients: a pilot study; de Lalla F et al.; Twenty-two non-neutropenic adult patients with Streptococcus pneumoniae meningitis received granulocyte-colony stimulating factor (G-CSF) (300-450 Ig/day subcutaneously for 6 days) in addition to cefotaxime plus dexamethasone (9-12 g/day for 10 days and 16 mg/day for 3 days iv, respectively) . Patients recovered without evident sequelae in all cases but one (with bilateral hearing deficit) . No adverse event was recorded . Improvement of inflammation indices in the cerebrospinal fluid was rapid . The most rapid improvement was seen in glucose concentration, which returned to normal ranges within 24-48 h of treatment . In this study G-CSF administration appeared to be safe and effective; further controlled clinical trials are justified.

J Antimicrob Chemother, 2000 Nov, 46(5), 767 - 73
Streptococcus pneumoniae resistance to erythromycin and penicillin in relation to macrolide and beta-lactam consumption in Spain (1979-1997); Granizo JJ et al.; Streptococcus pneumoniae resistance to penicillin and erythromycin in relation to beta-lactam and macrolide consumption in Spain over 19 years (1979-1997) was studied from resistance data collected by a search of the literature . Antibiotic consumption was expressed in defined daily dosage (DDD)/1000 inhabitants/day . A significant relationship (P: < 0.001) between erythromycin resistance (MIC >/= 1 mg/L) and global macrolide consumption (r = 0.942), as well as between high-level penicillin resistance (MIC >/= 2 mg/L) and global beta-lactam consumption (r = 0.948) was observed . The relationship between erythromycin resistance and macrolide consumption was due mainly to consumption of macrolides taken twice a day (adjusted r(2) = 0.886) . Prevalence of high-level penicillin resistance correlated with consumption of oral cephalosporins (adjusted r(2) = 0.877); however, there appeared to be no correlation of consumption of oral or parenteral aminopenicillins, narrow-spectrum penicillins or cephalosporins with intermediate-level penicillin resistance (MIC 0 . 12-1 mg/L) . The prevalence of high-level penicillin and of erythromycin resistance were also strongly correlated with each other (r = 0.903, P: < 0.001) . In addition to global consumption, different categories of resistance (high or intermediate), and the differential capability of antibiotics to select resistance, must be taken into account when studying antibiotic impact on bacterial populations . Although this ecological analysis is not able to demonstrate a causal relationship between antibiotic consumption and development of resistance, it suggests that overuse of certain specific antibiotics is more likely to be related to the increase in drug-resistant strains of S . pneumoniae.

J Antimicrob Chemother, 2000 Nov, 46(5), 757 - 65
Molecular epidemiology of penicillin-resistant Streptococcus pneumoniae colonizing children with community-acquired pneumonia and children attending day-care centres in Fortaleza, Brazil; Wolf B et al.; To study clonal diversity of penicillin-resistant Streptococcus pneumoniae, 161 randomly selected isolates with reduced susceptibility to penicillin, collected from the nasopharynx of children under 5 years of age with community-acquired pneumonia and healthy controls from public day-care and immunization centres in Fortaleza, Brazil, were characterized by microbiological and serological techniques and automated ribotyping . Also included were 44 randomly selected penicillin-susceptible strains and three international reference strains . With automated ribotyping 75 ribopatterns were observed: 50 ribogroups were unique and 25 ribogroups were represented by two or more isolates . Genetic diversity was extensive but some degree of genetic homogeneity was found in strains from children with pneumonia, strains from children in day-care centres, isolates with reduced susceptibility to penicillin and isolates expressing 'paediatric' serogroups . Fourteen (56%) clusters contained both isolates with reduced penicillin susceptibility and penicillin-susceptible isolates, suggesting emergence of penicillin resistance . In general, there was a good correlation between ribogroups and serogroups, but 12 (48%) clusters contained isolates with alternative serogroups . Isolates with such alternative serogroups were more often encountered in penicillin-susceptible strains (41%) than in strains with reduced susceptibility to penicillin (7%) . Thirty-eight (19%) isolates (including seven penicillin-susceptible strains) showed ribotypes indistinguishable from those of two international epidemic clones of S . pneumoniae: ribogroup 54-S-1 (15 isolates) with a ribopattern characteristic of the 23F multiresistant 'Spanish/USA' clone and ribogroup 74-S-3 (23 isolates) with a pattern similar to that of the 6B multiresistant 'Spanish' clone.

J Antimicrob Chemother, 2000 Nov, 46(5), 717 - 23
Subinhibitory concentrations of erythromycin reduce pneumococcal adherence to respiratory epithelial cells in vitro; Lagrou K et al.; We have investigated the influence of subinhibitory concentrations of erythromycin on the interaction between Streptococcus pneumoniae and human respiratory epithelial cells . Confluent in vitro cell cultures were inoculated with erythromycin-resistant S . pneumoniae and incubated for 24 h . Erythromycin significantly reduced adherence of the pneumococci after 4 h and 24 h: 4.0% +/- 0.7% (mean +/- S.E.M . ) of the pneumococci adhered to the epithelial cells in medium with erythromycin, compared with 7.7% +/- 0.8% in medium without erythromycin (P: = 0.002) after 4 h, and the corresponding values after 24 h were 24.2% +/- 5.3% and 38.4% +/- 5.0%, respectively (P: = 0.038) . Disruption of epithelial integrity by S . pneumoniae, measured as the decrease in transepithelial electrical resistance, was delayed in the presence of erythromycin . Neither addition of erythromycin to the culture medium nor infection of the cell cultures with pneumococci significantly affected secretion of interleukin-8 by the epithelial cells . Addition of erythromycin to a pneumococcal suspension in cell culture medium without respiratory epithelial cells almost completely prevented the release of pneumolysin . We conclude that erythromycin at subinhibitory concentrations reduces the adherence to and disruption of respiratory epithelial cells by S . pneumoniae, possibly by interfering with pneumolysin release.

J Antimicrob Chemother, 2000 Nov, 46(5), 669 - 83
Application of fluoroquinolone pharmacodynamics; Wright DH et al.; Pharmacodynamics provides a rational basis for optimizing dosing regimens by describing the relationship between drug, host and antimicrobial effect . The successful identification of meaningful pharmacodynamic outcome parameters can, therefore, greatly assist clinicians in making objective prescribing decisions rather than relying on static in vitro MIC data . While pharmacodynamic outcome parameters have been proposed for select antimicrobial agents, their clinical application remains to be defined fully . Quinolone antibiotics are generally considered to have concentration-dependent bactericidal activity and peak/MIC and AUC/MIC ratios have been identified as possible pharmacodynamic predictors of clinical and microbiological outcome as well as the development of bacterial resistance . Investigators have suggested that AUC/MIC ratios of 100-125 or peak/MIC ratios of >10 are required to predict clinical and microbiological success and to limit the development of bacterial resistance . These conclusions are derived primarily from studies of Gram-negative bacteria, and recent data suggest that these ratios may not be applicable for Streptococcus pneumoniae, where an AUC/MIC ratio of <40 appears to be a more accurate predictor . There is considerable variation in pharmacodynamic calculations and outcome parameters appear to be quinolone- and pathogen-specific . Additional prospective clinical research is needed to characterize quinolone pharmacodynamic parameters and answer unresolved questions regarding optimal pharmacodynamic outcome predictors for Gram-positive bacteria, anaerobes and atypical respiratory pathogens.

Pediatrics . 2000 Nov;106(5):E61.
Pneumococcal facial cellulitis in children; Givner LB et al.; OBJECTIVE: To review the epidemiology and clinical course of facial cellulitis attributable to Streptococcus pneumoniae in children . DESIGN: Cases were reviewed retrospectively at 8 children's hospitals in the United States for the period of September 1993 through December 1998 . RESULTS: We identified 52 cases of pneumococcal facial cellulitis (45 periorbital and 7 buccal) . Ninety-two percent of patients were <36 months old . Most were previously healthy; among the 6 with underlying disease were the only 2 patients with bilateral facial cellulitis . Fever (temperature: >/=100.5 degrees F) and leukocytosis (white blood cell count: >15 000/mm(3)) were noted at presentation in 78% and 82%, respectively . Two of 15 patients who underwent lumbar puncture had cerebrospinal fluid with mild pleocytosis, which was culture-negative . All patients had blood cultures positive for S pneumoniae . Serotypes 14 and 6B accounted for 53% and 27% of isolates, respectively . Overall, 16% and 4% were nonsusceptible to penicillin and ceftriaxone, respectively . Such isolates did not seem to cause disease that was either more severe or more refractory to therapy than that attributable to penicillin-susceptible isolates . Overall, the patients did well; one third were treated as outpatients . CONCLUSIONS: Pneumococcal facial cellulitis occurs primarily in young children (<36 months of age) who are at risk for pneumococcal bacteremia . They present with fever and leukocytosis . Response to therapy is generally good in those with disease attributable to penicillin-susceptible or -nonsusceptible S pneumoniae . Ninety-six percent of the serotypes causing facial cellulitis in this series are included in the heptavalent-conjugated pneumococcal vaccine recently licensed in the United States.

Rev Invest Clin, 2000 Jul-Aug, 52(4), 418 - 26
{Cost-effectiveness analysis of ceftriaxone and cefotaxime in the treatment of community-acquired pneumonia}; Garcia-Contreras F et al.; OBJECTIVE: To assess the cost-effectiveness ratio of ceftriaxone and cefotaxime to treat moderate to severe community acquired pneumonia (CAP) . METHODS: A clinical trial was done in five hospitals of the Instituto Mexicano del Seguro Social, at the metropolitan area of Mexico City . Ceftriaxone and cefotaxime were compared to treat moderate to severe CAP, and the costs of purchasing, preparation, administration, hospitalization, and therapeutic success were quantified . Cost-effectiveness ratio was calculated, and sensitivity analysis and incremental analysis were done . RESULTS: The main isolated germs were Streptococcus pneumoniae (23.6%) and Staphylococcus aureus (18.5%) . Most of the microorganisms were sensitive to ceftriaxone, ceftazidime, and cefotaxime, and were resistant to penicillin, ampicillin, and erythromycin . Therapeutic success was 98% in the ceftriaxone group and 83% in the cefotaxime group (p = 0.0091) . Cost-effectiveness ratio for per cent unit of success was $19,458.62 Mexican pesos in the ceftriaxone group and $29,218.08 in the cefotaxime group . Sensitivity analysis showed consistently a lower cost-effectiveness ratio in the ceftriaxone group . Incremental analysis based on the treatment of 55 patients showed that using ceftriaxone instead of cefotaxime resulted in saving $35,170.79 per each additional cured patient . CONCLUSIONS: Ceftriaxone has a lower cost-effectiveness ratio than cefotaxime to treat patients with CAP and bad prognosis criteria requiring hospitalization.

J Formos Med Assoc, 2000 Oct, 99(10), 783 - 4
Infected chylothorax caused by Streptococcus agalactiae: a case report; Wang JT et al.; Chylothorax is bacteriostatic in nature . Bacterial infection rarely develops in chylothorax and has never been reported in a non-immunocompromised host . A 33-year-old woman was admitted to National Taiwan University Hospital because of fever and right pleuritic pain . Chest roentgenography and computed tomography revealed right pleural effusion . Examination of the pleural effusion revealed a profile compatible with empyema and chylothorax . Culture of the pleural effusion yielded Streptococcus agalactiae . The woman was not immunocompromised . This is the first report of infected chylothorax caused by Streptococcus agalactiae in a non-immunocompromised host.

Expert Opin Investig Drugs, 2000 Aug, 9(8), 1711 - 21
Anti-adhesive strategies in the prevention of infectious disease at mucosal surfaces; Kelly CG et al.; Binding of microbial cell surface adhesins to host receptor molecules is a critical early step in microbial infection and pathogenesis . Anti-adhesive strategies aimed at blocking this interaction offer an attractive means of preventing infection at an early stage . The strategy should reduce the likelihood of resistant strains of microorganisms emerging, since those that do not bind will not be subjected to sustained selective pressure, as may occur with antibiotic therapy . Three classes of adhesion-blocking agent have been investigated, namely anti-adhesin antibodies, adhesin analogues and receptor analogues . The effectiveness of a number of these adhesion-blocking compounds has been demonstrated in human and animal models of infection . Direct application to the tooth surface of anti-adhesin monoclonal antibody, or a synthetic peptide adhesion epitope, prevented infection with the oral pathogen, Streptococcus mutans in humans . Intranasal administration of a soluble receptor analogue significantly reduced virus production and symptoms following experimental infection with rhinovirus . Similarly, all three types of anti-adhesion agent protected against a variety of infections at other mucosal surfaces in animal models . A common finding from these studies is the long duration of protection, which cannot be due to persistence of the anti-adhesion agent, but may be the result of competitive exclusion by members of the normal flora at specific mucosal surfaces . Development of these novel antimicrobial agents is particularly timely in view of the increasing concern over the spread of antibiotic resistance.

Adolesc Med, 2000 Oct, 11(3), 681 - 95
Community-acquired pneumonia in adolescents; Gordon RC; Community acquired pneumonia (CAP) is defined as pneumonia acquired outside of the hospital setting . Extensive studies of CAP in adolescents that characterize the true incidence of various etiologic pathogens are not available . However, Streptococcus pneumoniae, Mycoplasma pneumoniae, and Chlamydia pneumoniae appear to be the most frequently encountered pathogens . These organisms often cause CAP in adults as well; other infections are noted as well, including Legionella . "Atypical pneumonia" refers to pneumonia not presenting with the usual clinical picture of pneumococcal infection (which includes high fever, productive cough, chills, and other "classic" features) . The term is frequently used in adolescents with CAP . However, this classification may not help in individual patients, who often show a high degree of variability in the clinical presentation of pneumonia; also it does not always predict microbial cause . There is currently a trend away from the concept of atypical pneumonia syndrome and more discussion of atypical pathogens as commonly causes of CAP . This article reviews recent literature on CAP with special emphasis on its diagnosis and management in adolescent patients.

J Clin Microbiol, 2000 Nov, 38(11), 4137 - 44
Genetic diversity and clonal patterns among antibiotic-susceptible and -resistant Streptococcus pneumoniae colonizing children: day care centers as autonomous epidemiological units; Sa-Leao R et al.; Characterization by antibiotype of the 1,096 Streptococcus pneumoniae recovered from 2,111 nasopharyngeal samples of children attending 16 day care centers (DCCs) in Lisbon, Portugal, and molecular typing of 413 drug-resistant pneumococci (DRPn) and 89 fully drug-susceptible pneumococci (DSPn) has allowed several conclusions . (i) There was an increase in the frequency of DRPn colonizing children in DCCs from 40% in 1996 to 45% in 1997 to 50% in 1998 . (ii) Drug resistance spread by cross-transmission of DRPn clones . A few (8 out of 57) DRPn clones were repeatedly isolated from a large number of children in several DCCs and during each period of surveillance, suggesting the epidemic nature of these clones, which included lineages representing internationally spread S . pneumoniae clones . (iii) Dissemination of resistance determinants among pneumococci colonizing the nasopharynx occurred . Association of identical pulsed-field gel electrophoresis patterns with diverse antibiotypes among pneumococci colonizing children suggests that the high prevalence of DRPn involves not only cross-transmission of resistant strains but also dispersal of resistance genes through recombinational mechanisms . (iv) DCCs are autonomous epidemiological units . Among the 413 DRPn, 57 different lineages were detected; these lineages were dispersed among the 16 DCCs to produce unique microbiological profiles for each of the DCCs . Higher genetic diversity and less sharing of clonal types were observed among the DSPn.

J Clin Microbiol, 2000 Nov, 38(11), 4126 - 30
Analysis of immunoreactivity to a Streptococcus equi subsp . zooepidemicus M-like protein To confirm an outbreak of poststreptococcal glomerulonephritis, and sequences of M-like proteins from isolates obtained from different host species; Nicholson ML et al.; The etiologic agent of a large 1998 outbreak of poststreptococcal acute glomerulonephritis (PSGN) in Nova Serrana, Brazil, was found likely to be a specific strain of Streptococcus equi subsp . zooepidemicus from contaminated cheese (S . Balter et al., Lancet 355:1776-1780, 2000) . In the present study, we used a serologic screen for a known surface-exposed virulence factor to confirm the epidemiologic findings . Using primers flanking a previously characterized M-like protein gene (J . F . Timoney et al., Infect . Immun . 63:1440-1445, 1995), we amplified and sequenced the M-like protein (designated Szp5058) gene and found it to be identical among four independent acute-phase PSGN patient isolates . Convalescent-phase sera from 33 of 44 patients in the PSGN outbreak were found to contain antibodies highly reactive to a purified Szp5058 fusion protein, compared with 1 of 17 control sera (P < 0 . 0001), suggesting that Szp5058 was expressed during infection and further implicating this strain as the cause of the PSGN outbreak . The predicted signal sequence and cell wall association motif of Szp5058 were highly conserved with the corresponding sequence from S . equi subsp . zooepidemicus SzpW60, while the predicted surface-exposed portions differed markedly between these two proteins . The 5' end of the szp5058 gene, including its variable region, was identical to the szp gene from another strain associated with a previous PSGN outbreak in England (M . Barham et al., Lancet i:945-948, 1983), and the corresponding szp sequence found from the Lancefield group C type strain isolated from a guinea pig . In addition, the hypervariable (HV) portion of szp5058 was identical to a previously published HV sequence from a horse isolate (J . A . Walker and J . F . Timoney, Am . J . Vet . Res . 59:1129-1133, 1998) . Three other strains of S . equi subsp . zooepidemicus, including another strain previously associated with a PSGN outbreak, were each found to contain a distinct szp gene . Two of these szp genes had HV regions identical to szp regions from isolates recovered from different host species.

Cancer Res, 2000 Oct 15, 60(20), 5740 - 6
CM101 treatment overrides tumor-induced immunoprivilege leading to apoptosis; Yakes FM et al.; CM101, a bacterial polysaccharide exotoxin produced by group B Streptococcus (GBS), also referred to as GBS toxin, has been shown to target pathological neovasculature and activate complement (C3), thereby inducing neovascularitis, infiltration of inflammatory cells, inhibition of tumor growth, and apoptosis in murine tumor models . Data from refractory cancer patients in a Phase I clinical trial with CM101 indicated a similar mechanism of tumor-targeted inflammation . To further our understanding of the mechanism of action of CM101 as an antitumor agent, we examined the role of the inflammatory response in inducing tumor apoptosis in a normal mouse and tumor-bearing mouse model . The i.v . infusion of CM101 into B16BL-6 melanoma tumor-bearing mice elevated p53 mRNA in circulating leukocytes as measured by reverse transcription-PCR, and immunohistochemistry demonstrated infiltration and sequestration of leukocytes . Whole tumor lysates from excised tumors exhibited an increase in binding to the murine p21(Waf1/Cip1) derived p53 DNA binding sequence compared with control whole tumor lysates, in which minimal or no DNA binding was observed . CM101 infusion led to elevated levels of Fas protein within the tumors as well as a decrease in the expression of fas ligand (fasL) . Furthermore, tumors were apoptotic as determined by terminal deoxynucleotidyl transferase-mediated nick end labeling and DNA fragmentation assays . Collectively, these data suggest that CM101 up-regulates p53 in tumor-infiltrating leukocytes, initiating a loss of tumor immunoprivilege and consequently rendering the tumor sensitive to Fas/fasL-mediated apoptosis . CM101 induced loss of tumor immunoprivilege through changes in the expression of leukocyte p53, tumor Fas and fasL coupled with neovascularitis and leukocyte infiltration, constitutes a plausible molecular pathway for tumor reduction observed in cancer patients.

Pediatr Int, 2000 Oct, 42(5), 552 - 6
Nasopharyngeal colonization with penicillin-resistant Streptococcus pneumoniae in Turkish children; Ciftci E et al.; BACKGROUND: Streptococcus pneumoniae is one of the major infectious agents observed in children . In spite of the fact that penicillin is preferred in the treatment of infections caused by S . pneumoniae, there has been a world-wide increase in the frequency of penicillin-resistant S . pneumoniae . METHODS: One hundred and fifty sick children with a clinical diagnosis of pneumonia, meningitis, acute otitis media, acute sinusitis and septicemia or bacteremia, and 150 healthy children without any infection were examined . Streptococcus pneumoniae, which were isolated from the nasopharynx, were analyzed with respect to penicillin susceptibility using the agar dilution method . RESULTS: The S . pneumoniae carriage rate was observed to be 43.3% in the group of sick children and 30.0% in the control group (P < 0.05) . The penicillin resistance of S . pneumoniae isolated from the nasopharynx was determined to be 35.4% from a total of 110 isolates, with an intermediate resistance of 32.7% and a high resistance of 2.7% . The penicillin resistance of S . pneumoniae carried in the nasopharynx was determined to be 41.5% in the group of sick children and 26.6% in the control group (P > 0.05) . Resistance rates of other antibiotics were determined as follows: cefotaxime 2.7%, erythromycin 19%, clarithromycin 5.4%, tetracycline 21.8%, trimethoprim-sulfamethoxazole 4.5% and rifampin 0% . CONCLUSIONS: Penicillin resistance of S . pneumoniae has recently become a problem in Turkey . Because of this, we require new strategies to limit the spread of drug-resistant S . pneumoniae.

Sheng Wu Gong Cheng Xue Bao, 2000 May, 16(3), 396 - 9
{Nutrition condition of hyaluronic acid fermentation with Streptococcus zooepidemicus}; Gao HJ et al.; Based on the analysis of metabolic pathway Streptococcous zooepidemicus for hyaluronic acid (HA) synthesis, nucleotide, especially uracil, was considered to be important to cell growth and metabolism . When 0.005 g.L-1 uracil added in the media in which yeast extract as complex nitrogen source, cell growth and HA production were increased by 32% and 34% respectively . From analysis of amino acid in fermentation process, it was show that arginine(Arg) was needed for cell metabolism, and concentration of free Arg maintained at 0 g.L-1 in fermentation process, which was proposed to limit cell growth and HA production . By shake-flask experiment HA concentration reached 0.510 g.L-1 when 0.06 g.L-1 Arg added, in the fermentation with 2.5 L fermentor, when uracil 0.005 g.L-1 and Arg 0.06 g.L-1 were added, the rate of cell growth increased, maximum of specific growth rate, concentration of HA and HA molecular weight reached 0.67 h-1, 5.2 g.L-1 and 2.15 x 10(6) Da from 0.54 h-1, 4.2 g.L-1, 2.0 x 10(6) Da, respectively.

Minn Med, 2000 Oct, 83(10), 53 - 8
Antibiotic bacterial resistance in ambulatory patients; Yawn BP et al.; BACKGROUND: This study evaluates trends in antibiotic resistance in patients who were treated in an ambulatory setting . METHODS: The authors compiled the data from all lower respiratory track(sputum) cultures collected from ambulatory patients who visited the Olmsted Medical Center and Mayo Clinic between 1985 and 1998 . Cultured organisms were identified, and Minimal Inhibitory Concentration (MIC) values were presented and categorized as susceptible, intermediate, or resistant based on the National Committee for Clinical Laboratory Standards (NCCLS) guidelines for MIC and antibiotic susceptibility . RESULTS: 4,297 potentially pathogenic organisms were obtained from sputum cultures for 1,921 patients . The most discernible changes in antibiotic resistance appeared to be in cultures positive for Pseudomonas aeruginosa . A trend toward increasing resistance of isolates of Streptococcus pneumoniae to beta-lactam drugs was observed in a portion of the population . An emerging intermediate susceptibility among isolates of Klebsiella pneumoniae and Pseudoumonas species was noted . CONCLUSIONS: Trends in antimicrobial resistance of respiratory pathogens from ambulatory patients are less clear than those from hospitalized patients, but must be monitored because of the high percentage of ambulatory patients who receive empirical therapies . Trends in intermediate susceptibility patterns may help reveal emerging antimicrobial resistance.

Can Respir J, 2000 Sep-Oct, 7(5), 371 - 82
Summary of Canadian guidelines for the initial management of community-acquired pneumonia: an evidence-based update by the Canadian Infectious Disease Society and the Canadian Thoracic Society; Mandell LA et al.; Community-acquired pneumonia (CAP) is a serious illness with a significant impact on individual patients and society as a whole . Over the past several years, there have been significant advances in our knowledge and understanding of the etiology of the disease, and an appreciation of problems such as mixed infections and increasing antimicrobial resistance . The development of additional fluoroquinolone agents with enhanced activity against Streptococcus pneumoniae has been important as well . It was decided that the time had come to update and modify the previous CAP guidelines, which were published in 1993 . The current guidelines represent a joint effort by the Canadian Infectious Disease Society and the Canadian Thoracic Society, and they address the etiology, diagnosis and initial management of CAP . The diagnostic section is based on the site of care, and the treatment section is organized according to whether one is dealing with outpatients, inpatients or nursing home patients.

Eur J Clin Microbiol Infect Dis, 2000 Sep, 19(9), 704 - 7
Surveillance of penicillin-resistant Streptococcus pneumoniae in one American metropolitan area, 1989-1998; Sahloul RT et al.; The emergence of invasive penicillin-resistant (PRSP) and multidrug-resistant (MDRP) Streptococcus pneumoniae was tracked from 1989 to 1998 in one medium-sized metropolitan area in the USA, encompassing western West Virginia, including Huntington, the only major city, and neighboring sections of Kentucky and Ohio . Capsular serotyping and antibiotic sensitivity tests were performed on 350 community-acquired isolates comprising 93.1% of all pneumococcal isolates identified . The incidence of PRSP increased from 3 to 10% during the 10 years of the study . Twenty-nine (22.1%) of 131 isolates of serotypes 6, 9, 14, 19, and 23 were PRSP (one-fourth were MDRP) and 1 (0.5%) of 219 other serotypes was PRSP (serotype 35) . Invasive PRSP occurred most frequently in young children and in adults aged 80 years and older, 8.9 and 10.9 cases per 100,000 persons, respectively.

Eur J Gastroenterol Hepatol, 2000 Oct, 12(10), 1141 - 5
Thrombolytic therapy in patients with portal vein thrombosis: case report and review of the literature; Schafer C et al.; A 29-year-old male patient with Crohn's disease of the terminal ileum and previous abdominal surgery was admitted because of severe abdominal pain and signs of bacterial sepsis . The diagnosis of portal vein thrombosis and multiple liver abscesses due to Streptococcus intermedius septicaemia was made and antibiotic therapy was instituted immediately . As high-dose heparin therapy was ineffective, urokinase was administered intravenously over a total of 7 days . Within 2 days, the patient's symptoms completely subsided . Colour duplex ultrasonography revealed complete recanalization of the main stem of the portal vein; the right branch of the portal vein, however, remained occluded . Other case reports on thrombolytic therapy in patients with portal vein thrombosis are reviewed.

Scand J Infect Dis, 2000, 32(5), 564 - 5
Pyopneumothorax: a complication of Streptococcus pyogenes pharyngitis; Khatib R et al.; A 20-y-old African-American female with Streptococcus pyogenes pharyngitis presented with tension pyopneumothorax . Her illness began with fever and sore throat that persisted for several days . She then developed a left neck swelling, followed by difficult swallowing and cough . Subsequently, she developed shortness of breath that became severe . On physical examination fever (39.2 degrees C), exudative pharyngitis, tenderness and swelling in the left anterior cervical area were noted . Chest X-ray revealed left side pneumothorax, air-fluid level and near-complete collapse of the left lung with displacement of the heart and trachea to the right . Computed tomography scan of the neck revealed swelling and enhancement of the sternocleidomastoid muscle with loculated fluid collection, inflammation in the left anterior medial neck displacing the trachea extending into the mediastinum and the left apex . Thoracentesis revealed purulent fluid; Gram stain showed Gram-positive cocci in chains; culture yielded pure growth of Streptococcus pyogenes . She was treated with high dose penicillin, several chest tubes and intra-pleural injections of streptokinase with gradual resolution . This complication has not been described previously in Streptococcus pyogenes pharyngitis.

Scand J Infect Dis, 2000, 32(5), 491 - 4
Streptococcus pneumoniae septic arthritis in adults; James PA et al.; Septic arthritis is a rarely reported manifestation of disease due to Streptococcus pneumoniae . We have reviewed our recent experience of this disease in 14 adult patients . Common features in patients with S . pneumoniae septic arthritis included advanced age (median=63 y), pre-existing joint disease (6/14), large joint disease (14/14), polyarthritis (6/14), and associated meningitis, pneumonia or both (6/14) . Two patients with septic arthritis and meningitis, and another with Down's syndrome and sleep apnoea, died during treatment . In the remaining 11 patients, treatment for at least 19 d, predominantly with intravenous benzyl penicillin, plus joint lavage, resulted in cure.

Acta Crystallogr D Biol Crystallogr, 2000 Nov, 56 ( Pt 11), 1452 - 5
Crystallization and preliminary X-ray diffraction analysis of a functional form of pneumolysin, a virulence factor from Streptococcus pneumoniae; Kelly SJ et al.; Pneumolysin is a virulence factor from Streptococcus pneumoniae, a Gram-positive bacterial pathogen which causes human infections with a severe impact on mortality and morbidity worldwide . The enzyme belongs to a group of cholesterol-dependent cytolysins and interacts with its cholesterol receptor on target cells, leading to pneumolysin insertion into target-cell membranes and subsequently to pore formation and cell lysis . Pneumolysin has been overexpressed, purified and crystallized for X-ray diffraction studies . Crystals have been obtained in the presence of cholesterol in an effort to produce a three-dimensional structure of pneumolysin in its fully functional form with the enzyme bound to its activator . This is the first report of the crystallization of a cholesterol-dependent cytolysin in the presence of bound cholesterol . The vapor-diffusion method using ammonium sulfate as a precipitation agent was used to grow crystals in the presence of n-octyl-beta-D-glucopyranoside and phosphatidylcholine . Crystals of this 53 kDa molecule complexed with cholesterol diffracted X-rays to 3.3 A . The crystal unit cell has parameters a = b = 191.45, c = 66.16 A, alpha = beta = 90.0, gamma = 120 degrees and belongs to the trigonal space group P3 . The determination of the three-dimensional structure of this pneumococcal cytolysin is in progress.

Semin Respir Infect, 2000 Sep, 15(3), 184 - 94
The epidemiology of respiratory tract infections; File TM; Respiratory tract infections (RTIs) are the most common, and potentially most severe, of infections treated by health care practitioners . Lower RTIs along with influenza, are the most common cause of death by infection in the United States . Risk factors for pneumonia and other respiratory tract infections include: extremes of age (very young and elderly), smoking, alcoholism, immunosuppression, and comorbid conditions . The microbial cause of RTIs vary depending on the infection (i.e., pneumonia compared with acute bacterial sinusitis), setting (i.e., community-acquired compared with nosocomial), and other factors . The causative pathogens associated with CAP have changed in prevalence over time . Although Streptococcus pneumoniae remains the most common causative pathogen, a number of newer pathogens, such as Chlamydia pneumoniae and sin nombre virus, have been recognized in recent years . The emerging antimicrobial resistance of respiratory pathogens (most notably S . pneumoniae) has also increased the challenge for appropriate management of RTI . An awareness of the epidemiology and cause of specific respiratory infections should optimize care.

Rev Med Liege, 2000 Aug, 55(8), 775 - 81
{Antibiotics in pregnancy: importance of rational utilization}; Langhendries JP et al.; It is now clear that antibiotic treatment in the antenatal period significantly does prolong pregnancy during conservative management of preterm premature rupture of membranes and reduces neonatal infectious diseases as well as neonatal-related morbidities . In the same way, prophylactic intrapartum antibiotherapy reduces the incidence of early-onset group B Streptococcus-induced sepsis . Nevertheless, on the other hand, antibiotics in the perinatal period are associated with an increase of neonatal sepsis by organisms resistant to maternally administered antibiotics . In addition, antibiotic treatment in this period of time is emerging as one of the possible sources of the dramatic increase in atopic disorders in infants and children owing to the interference with the normal process of intestinal microbial colonization . So, guidelines for using antibiotics in the perinatal period can be said as one of the major priority in public health . Antibiotics have therefore to be rightly choosen and must be used in a rational manner . Local microbial epidemiology, period of infection onset, clinical evaluation, all together allow the physician to use antibiotics, always in association, according to the "well-thought-out wager" . In addition, the pharmacodynamic/pharmacokinetic relationship of each drug has to be known, in order to increase efficacy, decrease toxicity and reduce microbial resistance . It is especially mandatory in neonatology where the differences in drug distribution and drug elimination are of great concern, as compared to children and adults . The aim of this paper is to point out such very important aspects using antibiotics in the perinatal period.

Vet Res, 2000 Sep-Oct, 31(5), 473 - 9
Production of muraminidase-released protein (MRP), extracellular factor (EF) and suilysin by field isolates of Streptococcus suis capsular types 2, 1/2, 9, 7 and 3 isolated from swine in France; Berthelot-Herault F et al.; A total of 323 isolates of Streptococcus suis recovered from diseased or healthy pigs in France were serotyped . The presence of virulence-related proteins, Muraminidase-Released Protein (MRP), Extracellular Factor (EF) and Suilysin was also studied in 122 isolates of capsular types 2, 1/2, 9, 7 and 3 to evaluate their implication in virulence of S . suis . Capsular types 2, 1/2, 9, 7 and 3 were the most frequently detected (93%), with 69% for the capsular type 2 alone . Capsular types 2, 1/2, 9, 7, 3, 1, 4, 8, 18, 10 and 12 were isolated from diseased pigs, whereas types 2, 7, 9, 1/2, and 3 originated from the nasal cavities or tonsils of healthy animals . Most of the S . suis type 2 isolates recovered from diseased pigs carried MRP+ EF- Suilysin- (46%) or MRP+ EF+ Suilysin+ (28%) phenotypes . The MRP+ EF- Suilysin- phenotype was also detected in 67% of S . suis type 2 strains isolated from healthy pigs . The production of the virulence-related proteins was less frequently found in S . suis types 1/2, 9, 7 and 3 recovered either from diseased or healthy pigs . In this study, all the capsular type 1/2 strains were MRP+ EF- Suilysin- and all the S . suis type 7 harboured an MRP- EF- Suilysin- phenotype . The MRP- EF- Suilysin- phenotype was found in S . suis types 2, 3, 7 and 9 isolated from septicaemia, meningitis, pneumonia, and pleurisy . These results suggest that the presence of these proteins should not be used as a single condition for classifying the virulence of a field isolate in France.

Curr Opin Microbiol, 2000 Oct, 3(5), 528 - 34
New agents for Gram-positive bacteria; Bhavnani SM et al.; Infections caused by multiple-resistant Gram-positive organisms continue to occur at an alarming rate worldwide . Two new and unique antimicrobial agents targeted specifically against such organisms, quinupristin/dalfopristin and linezolid, have been approved for use in the USA in the past year and will play an important role in the treatment of life-threatening infections . In addition, several new fluoroquinolones have been approved recently or will be available in the near future to aid in the treatment of infections caused by resistant strains of Streptococcus pneumoniae.

Curr Opin Microbiol, 2000 Oct, 3(5), 502 - 7
Pneumococcal conjugate vaccines - implications for community antibiotic prescribing; Tan TQ; Infections caused by Streptococcus pneumoniae are a major cause of morbidity and mortality in the pediatric population worldwide . Development of increasing resistance to multiple classes of antibiotics is making treatment of infections caused by this organism much more difficult . In order to prevent disease, a 23-valent pneumococal polysaccharide vaccine is available . However, this vaccine is poorly immunogenic in infants and young children . The development and licensing of pneumococcal conjugate vaccines that are safe and effective in the pediatric population is an important step in our ability to decrease the prevalence of pneumococcal disease seen.

Clin Infect Dis, 2000 Oct, 31(4), E15 - 7 Epub 2000 Oct 17.
Report of 2 fatal cases of adult necrotizing fasciitis and toxic shock syndrome caused by Streptococcus agalactiae; Tang WM et al.; We describe 2 cases of fatal necrotizing fasciitis and toxic shock syndrome caused by Streptococcus agalactiae-a rare entity that has been reported in only 9 patients-in 2 nonpregnant adults.

Clin Infect Dis, 2000 Oct, 31(4), 1008 - 11 Epub 2000 Oct 13.
Breakthrough pneumococcal bacteremia in patients being treated with azithromycin and clarithromycin; Kelley MA et al.; Streptococcus pneumoniae strains have exhibited decreasing susceptibility to penicillins and macrolides during the past several years . We reviewed the medical charts of all patients with pneumococcal bacteremia who were admitted to a university hospital over a period of 1 year, to identify failures of outpatient therapy . Of 41 patients admitted with pneumococcal bacteremia, 4 had previously taken either azithromycin or clarithromycin for 3-5 days . All 4 had pneumococcal strains that exhibited low-level resistance to macrolide antibiotics . Among pneumococci, low-level resistance to macrolides can lead to clinical failure, and resistance to macrolides should be considered during the selection of empiric therapy for patients with presumed pneumococcal infections.

Clin Infect Dis, 2000 Oct, 31(4), 887 - 93 Epub 2000 Oct 25.
Antimicrobial use and colonization with erythromycin-resistant Streptococcus pneumoniae in Greece during the first 2 years of life; Syrogiannopoulos GA et al.; We evaluated nasopharyngeal colonization with erythromycin-resistant Streptococcus pneumoniae during the first 2 years of life in central and southern Greece . Of 2448 children studied from February 1997 to February 1999, 766 (31%) carried 781 pneumococcal isolates . Ninety-five (3.9%) of the children attended day care centers . Eighteen percent of the pneumococci were resistant to erythromycin (minimal inhibitory concentration 1 to >128 microg/mL), with 67.9% of them carrying the erm(B) gene and 29.2% mef(A) gene products . Four strains possessed neither the erm(B) nor the mef(A) gene . Multidrug resistance occurred in 97% and 40% of isolates carrying the erm(B) and mef(A) gene, respectively . An association was found between the erm(B) gene and serotypes 6B and 23F and between the mef(A) gene and serotypes 14 and 19F . A significant relationship existed between carriage of erythromycin-resistant pneumococci and use of macrolides or beta-lactams in the previous 3 months; the association was strongest when macrolide therapy was administered during the last month (odds ratio, 5.92; P=.0001) . The findings indicate the necessity of a judicious use of both macrolides and beta-lactams in young children to reduce the colonization with erythromycin-resistant pneumococci and the subsequent spread of such strains to the community.

Ned Tijdschr Geneeskd, 2000 Oct 7, 144(41), 1937 - 41
{Influenza A pneumonia}; Veenstra RP et al.; The majority of influenza cases are not associated with complications . Secondary bacterial pneumonia, commonly caused by Streptococcus pneumoniae or Staphylococcus aureus, is well known to most clinicians . Primary influenza viral pneumonia, characterized by rapidly progressive hypoxia and respiratory insufficiency together with non-consolidating pulmonary infiltrates, has a high mortality rate . In 3 patients, a man aged 74 years, and two neonates aged 11 months and 4 weeks respectively, primary influenza A pneumonia was diagnosed . In the latter two patients the virus was cultivated from sputum . Despite intensive supporting and drug treatment, the first and the last patients died . In view of evolving therapeutic possibilities, notably regarding neuraminidase inhibitors, it is important that clinicians recognize this complication of influenza at an early stage.

Lijec Vjesn, 2000 Jul-Aug, 122(7-8), 160 - 4
{Monitoring bacterial resistance to antibiotics in the CroatianRepublic}; Tambic T et al.; In 1996 a Committee for antibiotic resistance surveillance in Croatia was founded by the Croatian Academy of Medical Sciences . In this study antibiotic surveillance results for the period June 1-December 31, 1997 from 12 microbiology laboratories throughout Croatia are presented . Sensitivity to antibiotics was determined by disk diffusion method for the following bacteria: Streptococcus pneumoniae, Staphylococcus aureus, Enterococcus spp., Escherichia coli, Klebsiella spp . and Pseudomonas aeruginosa . In general, high proportion of resistant isolates was recorded throughout Croatia, although some regional variations were noticed . Mean resistance of pneumococci to penicillin was 38%, in S . aureus resistance to methicillin was 47%, and 3rd generation cephalosporin-resistance in E . coli was 6% and in Klebsiella spp . 21% . In P . aeruginosa resistance to gentamicin averaged 50%, to imipenem 13% and to ceftazidim 8% . Future aims of the Committee are to continue routine antibiotic resistance surveillance during certain periods every year, and to estimate clinical significance of resistant bacteria, detect mechanisms of resistance and improve the quality of laboratory work through education and quality control projects.

Protein Sci, 2000 Sep, 9(9), 1847 - 51
Structure of streptococcal pyrogenic exotoxin A reveals a novel metal cluster; Earhart CA et al.; The streptococcal pyrogenic toxins A, B, and C (SPEA, SPEB, and SPEC) are responsible for the fever, rash, and other toxicities associated with scarlet fever and streptococcal toxic shock syndrome . This role, together with the ubiquity of diseases caused by Streptococcus pyogenes, have prompted structural analyses of SPEA by several groups . Papageorgiou et al . (1999) have recently reported the structure of SPEA crystallized in the absence of zinc . Zinc has been shown to be important in the ability of some staphylococcal and streptococcal toxins to stimulate proliferation of CD4+ T-cells . Since cadmium is more electron dense than zinc and typically binds interchangeably, we grew crystals in the presence of 10 mM CdCl2 . Crystals have been obtained in three space groups, and the structure in the P2(1)2(1)2(1) crystal form has been refined to 1.9 A resolution . The structural analysis revealed an identical tetramer as well as a novel tetrahedral cluster of cadmium in all three crystal forms on a disulfide loop encompassing residues 87-98 . No cadmium was bound at the site homologous to the zinc site in staphylococcal enterotoxins C (SECs) despite the high structural homology between SPEA and SECs . Subsequent soaking of crystals grown in the presence of cadmium in 10 mM ZnCl2 showed that zinc binds in this site (indicating it can discriminate between zinc and cadmium ions) using the three ligands (Asp77, His106, and His110) homologous to the SECs plus a fourth ligand (Glu33).

J Microbiol Immunol Infect, 2000 Sep, 33(3), 187 - 90
Clinical presentation of acute mastoiditis in children; Jiang CB et al.; Nineteen children with 21 episodes of acute mastoiditis were treated in our hospital from 1989 to 1998 . The diagnosis was based on physical, radiologic, and surgical findings . The affected children were aged from 1 year old to 17 years old, with the peak incidence at 4 years old (23.8%) . Postauricular pain (90.5%) and fever (81%) were the most common harbingers of incipient acute mastoiditis . Streptococcus pneumoniae (38.1%) was the most common organism isolated followed by Pseudomonas aeruginosa (23.8%) . Underlying diseases such as leukemia and myeloid metaplasia were found in 38.6% of patients . All of the patients were initially treated with intravenous antibiotics during hospitalization . Six patients were managed with an adjunctive drainage procedure such as myringotomy or mastoidectomy . The most common complication of acute mastoiditis was hearing loss (31.6%); the second was meningitis (21.1%) . Subperiosteal abscess was found in two patients and brain abscess in one . Although acute mastoiditis is an uncommon condition, early diagnosis and management are necessary to prevent more serious complications.

Graefes Arch Clin Exp Ophthalmol, 2000 Sep, 238(9), 797 - 800
Ligneous conjunctivitis in a girl with severe type I plasminogen deficiency; Kraft J et al.; BACKGROUND: Ligneous conjunctivitis is a rare form of chronic recurrent pseudomembranous disease and may be associated with systemic membranous pathological changes . Recently ligneous conjunctivitis has been linked to severe type I plasminogen deficiency . We report on a patient with plasminogen deficiency and severe bilateral ligneous conjunctivitis . A new treatment approach and its outcome in this patient are described . CASE REPORT: We present the case of a 9-month-old Turkish girl with massive swelling of the eyelids and hard white pseudomembranes on both lids . The conjunctival smear was positive for Streptococcus pneumoniae . The clinical diagnosis was: ligneous conjunctivitis with superinfection . Histological investigation showed fibrin as major component of the pseudomembranes . The coagulation analyses revealed decreased plasminogen activity (<5%; normal 80-120%) and decreased plasminogen antigen (<0.4 mg/dl; normal 6-25 mg/dl) . The failure of surgical therapy led to the attempt at treatment with intravenous lys-plasminogen . A significant improvement of the ocular symptoms occurred; stabilization with no recurrent pseudomembranes could be achieved for 6 months after treatment . DISCUSSION: The initial amelioration of symptoms in our patient after systemic replacement therapy confirms the etiological importance of plasminogen deficiency in the development of ligneous conjunctivitis . Curative treatment of ligneous conjunctivitis is still not available . However, intravenous application of plasminogen offers new possibilities in therapy, although long-term treatment seems necessary.

Int J Dermatol, 2000 Sep, 39(9), 678 - 88
The significance of stress hormones (glucocorticoids, catecholamines) for eruptions and spontaneous remission phases in psoriasis; Weigl BA; BACKGROUND: In an earlier paper, it was described how acute eruptions of psoriasis may be produced in phases of immune deficiency and in the presence of bacterial antigen-releasing inflammatory foci, whereas clinical spontaneous remissions are produced in phases of immunologic activity . Therefore, it was of interest to investigate whether the stress hormones cortisol/epinephrine are involved in triggering such deficiency and activity phases . METHODS: During a series of investigations lasting up to 3 years in 95 patients, the following were determined: cortisol/epinephrine levels, polyclonal serum immunoglobulins IgM, IgG, and IgA, total serum IgE, complement C3 and C4 proteins, T cells and subpopulations, as well as streptococcal titers ASO/ADNase B, severity index (PASI) RESULTS: Phases of clinical inactivity are associated with the mechanism, "immunologic regulation," where antibacterial titers are elevated, but all other parameters are unremarkable . Eruption phases (in 32 of 95 patients) showed absolute increases in serum cortisol levels and antibacterial titers, and decreases in serum epinephrine (adrenaline) levels . Phases of spontaneous remission (in 25 of 32 patients) showed, in contrast to the eruption phases, absolute increases in serum epinephrine levels, and significant falls in serum cortisol levels and bacterial titers . CONCLUSIONS: On the basis of these results, the participation of the immune system is confirmed in the pathogenesis of psoriasis, which is subject to control by higher neurohormonal systems . Cortisol may be involved in the clinical eruption phase, and epinephrine in the remission phase . Both hormones are true antagonists and have important effects on the human immune system if produced in excess via the pituitary-adrenal axis . Infection with Streptococcus pyogenes is an additional trigger for the dermatosis.

WMJ, 2000 Aug, 99(5), 55 - 9
Antibiotic susceptibility of invasive Streptococcus pneumoniae in Wisconsin, 1999; Belongia EA et al.; BACKGROUND: Streptococcus pneumoniae is a major cause of community acquired infections in the United States, and rates of antibiotic resistance have increased dramatically in the past decade . Statewide rates of pneumococcal resistance to penicillin and other antibiotics have not been previously reported in Wisconsin . To determine these rates, we assessed invasive pneumococcal isolates for reduced susceptibility to nine different antibiotics . METHODS: Pneumococcal isolates from blood, cerebrospinal fluid or other normally sterile body sites were submitted by 91% of laboratories that perform invasive bacterial cultures . Isolates were tested for susceptibility to penicillin, cefotaxime, ceftriaxone, levofloxacin, meropenem, erythromycin, vancomycin, sulfamethoxazole-trimethoprim and chloramphenicol . RESULTS: There were 409 invasive pneumococcal isolates identified in 1999 among Wisconsin residents, including 385 (94%) isolates from blood . The mean patient age was 42.5 years (range, < 1 year to 96 years), and 213 (52%) were male . Of the pneumococcal isolates, 24% were not susceptible to penicillin, including 10% with high level resistance . Isolates with reduced penicillin susceptibility were also likely to have reduced susceptibility to other antibiotics . Patients with penicillin nonsusceptible (intermediate and fully resistant) pneumococcal isolates were significantly younger (mean, 37.0 years) than those with susceptible isolates (mean, 44.3 years) (p = .04) . The proportion of patients with a penicillin nonsusceptible isolate varied by region, ranging from 12.8% in northeastern Wisconsin to 35.5% in northern Wisconsin . CONCLUSIONS: The proportion of invasive pneumococcal isolates with penicillin resistance in Wisconsin is similar to other regions of the United States . Inappropriate antibiotic use contributes to the emergence of resistant pneumococcal infections, and educational efforts are underway to promote judicious antibiotic use in Wisconsin.

J Pak Med Assoc, 2000 Sep, 50(9), 289 - 93
The efficacy and safety of cefaclor in respiratory infections amongst Pakistani children; Najam Y et al.; PURPOSE: To evaluate the efficacy and safety of Cefaclor in respiratory tract infections amongst Pakistani children . PATIENTS AND METHODS: Multicenter, open label and non-comparative study was done to evaluate the response in terms of symptoms (In vivo) and bacterial cultures (In Vitro) to Cefaclor amongst children with respiratory tract infection between the ages 2 months to 12 years . Each patient was asked to visit the doctor on three occasions i.e., Day 0 (Initial evaluation prior to commencement of study), Day 4 (During therapy assessment and confirmation of compliance) and Day 10 (End of therapy assessment and compliance evaluation) . Representative swab specimens (Throat swabs, Ear swabs or Sputum) were collected from the infected site on day 0 and day 10 for culture and sensitivity . Patients were also assessed by the evaluators on each visit in terms of clinical symptomatic response and information collected was documented on a prescribed data base form . RESULTS: A total of 160 patients were enrolled in the study, of whom 15 were lost to follow-up between the first and second visit and a further 38 were lost by the 3rd visit . Thus 107 patients completed the study as per protocol . Otitis media and Upper respiratory tract infection were the predominant ailments amongst the cases enrolled . One or more bacteria were isolated in 75 (46%) instances, the maximum number of isolates being from ear swabs of Otitis media patients . Beta haemolytic Streptococcus (group A,C,F,G) seen in 18 cases was the most common pathogen reported followed by Staphylococcus aureus, H . influenzae and Streptococcus pneumoniae in 13,12 and 11 cases respectively . Sensitivity of Cefaclor for bacteria commonly seen in the respiratory tract was greater than 90% in most of the cases . Evaluation of the 42 culture proven cases for patients who completed the study showed that Cefaclor had a 93% efficacy for indicated bacteria and 54% for non-indicated bacteria . In Vivo analysis of Cefaclor (i.e . on the basis of symptomatic response) showed that 96% cases had a symptomatic response by the second visit, which improved to 97% by the third visit . Only 15 non-serious adverse events were observed in 160 patients, none of the cases necessitated discontinuation of drug . Mild gastrointestinal symptom was the most common adverse event reported . CONCLUSION: Cefaclor was found to be a safe and efficacious drug in the treatment of bacterial respiratory tract infections amongst Pakistani children.

Mayo Clin Proc, 2000 Oct, 75(10), 1008 - 14
Infective endocarditis in patients receiving long-term hemodialysis; McCarthy JT et al.; OBJECTIVE: To ascertain the predominant characteristics of patients receiving long-term dialysis who develop infective endocarditis (IE) . PATIENTS AND METHODS: We reviewed the records of all chronic hemodialysis patients who had IE at Mayo Clinic, Rochester, Minn, between 1983 and 1997 . RESULTS: Twenty episodes of IE occurred in 17 patients . One patient had 3 episodes of IE, and 1 patient had 2 episodes of IE; each episode was caused by a different organism . The mean +/- SD age of our patients was 63 +/- 11 years; there were 13 males; 6 patients had diabetes mellitus; and the mean +/- SD duration of hemodialysis prior to IE was 24.2 +/- 20.5 months . This analysis included 10 episodes of IE (occurring in 9 patients) within the Mayo Clinic Dialysis System during which time 223,358 hemodialysis treatments were delivered, giving a rate of 10 IE episode per 223,336 hemodialysis treatments . Among all 20 IE episodes, there were 14 synthetic arteriovenous grafts, 4 permanent venous dialysis catheters, 2 temporary venous dialysis catheters, and 2 native arteriovenous fistulas (2 accesses in 2 patients), and access had been in place for a mean +/- SD of 15.9 +/- 18.6 months . The portal of infection was the hemodialysis access in 13 episodes of IE . The causative organisms for IE were Staphylococcus aureus in 8 cases, Enterococcus sp in 4 cases, viridans streptococcus in 3 cases, Staphylococcus epidermidis in 2 cases, and 1 case each of Streptococcus bovis, group G beta-hemolytic streptococcus, and Aspergillus sp . The mitral valve was involved in 9 cases, the aortic valve was involved in 5 cases, and the tricuspid and pulmonic valves were involved in 1 case each . Patient survival (after the first episode of IE) was 71% at 30 days; 53% at 60 days; and 35% at 1 year . Echocardiography was performed in 19 episodes of IE . The transthoracic echocardiogram was 62.5% sensitive and 40% specific for the presence of definite or probable vegetations . Univariate analysis for factors affecting 60-day survival show that presence of right-sided IE, vegetation size greater than 2.0 cm3, diagnosis of diabetes mellitus, and initial leukocyte count greater than 12.5 x 10(9)/L were poor prognostic factors . Aortic valve involvement carried a better prognosis . CONCLUSIONS: Infective endocarditis in hemodialysis patients is relatively infrequent but has a high mortality . Patients with synthetic intravascular dialysis angioaccess (synthetic grafts and venous catheters) are more likely to develop IE than patients with native arteriovenous fistulas . Transesophageal echocardiography is a preferred echocardiographic study for suspected cases of IE . Prolonged antibiotic therapy is needed for all patients, and close monitoring is needed for patients with right-sided IE, large vegetations, diabetes mellitus, and an elevated leukocyte count.

Arch Dis Child, 2000 Nov, 83(5), 413 - 4
Streptococcus pneumoniae and Mycoplasma pneumoniae coinfection in community acquired pneumonia; Toikka P et al.; The characteristics of nine children with community acquired pneumonia with evidence of Streptococcus pneumoniae and Mycoplasma pneumoniae coinfection are described.

Arch Dis Child, 2000 Nov, 83(5), 408 - 12
Community acquired pneumonia--a prospective UK study; Drummond P et al.; BACKGROUND: There are few data on paediatric community acquired pneumonia (PCAP) in the UK . AIMS: To investigate the aetiology and most useful diagnostic tests for PCAP in the north east of England . METHODS: A prospective study of hospital admissions with a diagnosis of PCAP . RESULTS: A pathogen was isolated from 60% (81/136) of cases, and considered a definite or probable cause of their pneumonia in 51% (70/136) . Fifty (37%) had a virus implicated (65% respiratory syncytial virus) and 19 (14%) a bacterium (7% group A streptococcus, 4% Streptococcus pneumoniae), with one mixed infection . Of a subgroup (51 patients) in whom serum antipneumolysin antibody testing was performed, 6% had evidence of pneumococcal infection, and all were under 2 years old . The best diagnostic yield was from paired serology (34%, 31/87), followed by viral immunofluorescence (33%, 32/98) . CONCLUSION: Viral infection accounted for 71% of the cases diagnosed . Group A streptococcus was the most common bacterial infective agent, with a low incidence of both Mycoplasma pneumoniae and S pneumoniae . Pneumococcal pneumonia was the most common bacterial cause of pneumonia in children under 2 years but not in older children . Inflammatory markers and chest x ray features did not differentiate viral from bacterial pneumonia; serology and viral immunofluorescence were the most useful diagnostic tests.

Medicine (Baltimore), 2000 Sep, 79(5), 327 - 37
Streptococcus pneumoniae endocarditis in adults . A multicenter study in France in the era of penicillin resistance (1991-1998) . The Pneumococcal Endocarditis Study Group; Lefort A et al.; To better define the overall characteristics and risk factors for dying of adult pneumococcal endocarditis (PE) focusing on the echocardiographic diagnosis, the impact of surgery, and emergence of penicillin resistance, the medical and microbiologic charts of adult PE cases observed between 1991 and 1998 in university and general hospitals were reviewed through a nationwide retrospective study in France . Thirty cases of PE (22 men, 8 women; median age, 53 yr; range, 27-87 yr) were collected and validated . Twenty patients (66.7%) had no known predisposing cardiopathy; 4 had a bioprosthetic valve . The primary focus of infection was pneumonia in 10 (33.3%), and meningitis was noted in 12 (40.0%) . Half the patients suffered from chronic alcoholism . Echocardiography detected vegetation(s) in 29 cases (96.7%), valvular perforation in 6 (20.0%), and/or valve ring abscess in 4 (13.3%) . The most frequent complications were congestive heart failure (n = 19), large arterial emboli (n = 8), and focal abscesses (n = 7) . Five strains were penicillin-resistant . Twenty (66.7%) patients underwent valve replacement, 12 of them during the first month . The overall mortality rate was 24.1% . According to a multivariate analysis, the risk factors independently associated with dying were age > or = 65 yr and septic shock, while cardiac surgery was protective (p < 0.01) . In conclusion, PE is usually fulminant and causes severe valve damage and embolic complications; its short-term prognosis might be improved by early valve replacement.

J Dent Res, 1988 Jan, 67(1), 21 - 3
The effect of desalivation on coronal and root surface caries in rats; Bowen WH et al.; Although the presence of coronal caries is declining in much of the Western Hemisphere, the prevalence of root surface caries is likely to increase as teeth are retained longer than heretofore . At the same time, an increasing number of the population suffer from dry mouth as a result of taking prescription drugs, with an apparent concomitant increased susceptibility to root surface caries . This study attempted to develop an animal model which would aid in the exploration of the effects of desalivation and the development of root surface caries . Animals were desalivated, infected with Actinomyces viscosus and Streptococcus mutans (sobrinus) 6715, and fed a cariogenic diet . Coronal caries developed rapidly in the animals; sufficient disease was present after two weeks to permit evaluation of potential therapeutic agents . Alveolar bone loss and root surface lesions developed in three to four weeks . S . mutans (sobrinus) and A . viscosus established readily in all animals; however, as the investigation progressed, populations of the latter declined, possibly because of the highly acidogenic environment . This model will facilitate investigation of the influence of hyposalivation and help in the exploration of agents to alleviate the adverse effects of salivary gland dysfunction.

Eur J Oral Sci, 2000 Oct, 108(5), 393 - 402
Automated immunofluorescence for enumeration of selected taxa in supragingival dental plaque; Gmur R et al.; The present study investigated a recently developed automated image analysis technique for its applicability to the enumeration of selected bacteria in supragingival dental plaque . Following initial calibration, the system is capable to count fluorescence-labeled target cells in up to 48 samples without user interference . Test samples contained a characteristic mixture of planktonic bacteria, small almost planar bacterial aggregates, and large, virtually indisruptable clumps with cells from multiple species . Due to their complex composition, these samples provided a challenging validation step for the image analysis system . Automated enumeration of target bacteria was compared with visual counting of the fluorescence-labeled bacteria . Results are shown for six taxa (Actinomyces naeslundii, Fusobacterium nucleatum, Prevotella intermedia/Prev . nigrescens, Streptococcus gordonii/Strep . oralis/Strep . sanguis, Strep . sobrinus, and Veillonella dispar/ V . parvula) with characteristic differences in abundance, cell morphology and aggregation behavior . Results revealed good correspondence between the two enumeration techniques (correlation coefficients ranging from 0.77 to 0.92) provided that the portion of target bacteria exceeded 0.05% of the total bacterial cell number . This work demonstrates the applicability and usefulness of fully automated immunofluorescence to analyze such complex ecosystems as supragingival dental plaque.

Acta Clin Belg, 2000 Jul-Aug, 55(4), 222 - 4
Streptococcus milleri, a rare cause of pericarditis; successful treatment by pericardiocentesis combined with parenteral antibiotics; Marchal LL et al.; We report the case of a 71-year-old woman suffering from purulent pericarditis, but displaying only minor symptoms . No predisposing factors were found . In addition, the infection was caused by Streptococcus milleri, a rare causative agent for this affection . We review the literature and discuss the different types of treatment of purulent pericarditis.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 3196 - 8
Resistance to multiple fluoroquinolones in a clinical isolate of Streptococcus pyogenes: identification of gyrA and parC and specification of point mutations associated with resistance; Yan SS et al.; A strain of Streptococcus pyogenes resistant to multiple fluoroquinolones was isolated from the blood of an immunocompromised patient . Resistance to fluoroquinolones in S . pyogenes has not been previously studied . Compared to 10 sensitive strains of S . pyogenes, the fluoroquinolone-resistant clinical isolate of S . pyogenes presented point mutations in gyrA, predicting that serine-81 was changed to phenylalanine and that methionine-99 was changed to leucine, and in parC, predicting that serine-79 was changed to tyrosine . The mechanism of fluoroquinolone resistance in this isolate of S . pyogenes appears to be analogous to previously reported mechanisms for Streptococcus pneumoniae.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 3112 - 7
Potent antipneumococcal activity of gemifloxacin is associated with dual targeting of gyrase and topoisomerase IV, an in vivo target preference for gyrase, and enhanced stabilization of cleavable complexes in vitro; Heaton VJ et al.; We investigated the roles of DNA gyrase and topoisomerase IV in determining the susceptibility of Streptococcus pneumoniae to gemifloxacin, a novel fluoroquinolone which is under development as an antipneumococcal drug . Gemifloxacin displayed potent activity against S . pneumoniae 7785 (MIC, 0.06 microgram/ml) compared with ciprofloxacin (MIC, 1 to 2 microgram/ml) . Complementary genetic and biochemical approaches revealed the following . (i) The gemifloxacin MICs for isogenic 7785 mutants bearing either parC or gyrA quinolone resistance mutations were marginally higher than wild type at 0.12 to 0.25 microgram/ml, whereas the presence of both mutations increased the MIC to 0.5 to 1 microgram/ml . These data suggest that both gyrase and topoisomerase IV contribute significantly as gemifloxacin targets in vivo . (ii) Gemifloxacin selected first-step gyrA mutants of S . pneumoniae 7785 (gemifloxacin MICs, 0.25 microgram/ml) encoding Ser-81 to Phe or Tyr, or Glu-85 to Lys mutations . These mutants were cross resistant to sparfloxacin (which targets gyrase) but not to ciprofloxacin (which targets topoisomerase IV) . Second-step mutants (gemifloxacin MICs, 1 microgram/ml) exhibited an alteration in parC resulting in changes of ParC hot spot Ser-79 to Phe or Tyr . Thus, gyrase appears to be the preferential in vivo target . (iii) Gemifloxacin was at least 10- to 20-fold more effective than ciprofloxacin in stabilizing a cleavable complex (the cytotoxic lesion) with either S . pneumoniae gyrase or topoisomerase IV enzyme in vitro . These data suggest that gemifloxacin is an enhanced affinity fluoroquinolone that acts against gyrase and topoisomerase IV in S . pneumoniae, with gyrase the preferred in vivo target . The marked potency of gemifloxacin against wild type and quinolone-resistant mutants may accrue from greater stabilization of cleavable complexes with the target enzymes.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 3101 - 6
Evernimicin (SCH27899) inhibits a novel ribosome target site: analysis of 23S ribosomal DNA mutants; Adrian PV et al.; Spontaneous mutants of susceptible clinical and laboratory isolates of Streptococcus pneumoniae exhibiting reduced susceptibility to evernimicin (SCH27899; MIC, 0.5 to 4.0 mg/liter) were selected on plates containing evernimicin . Four isolates that did not harbor mutations in rplP (which encodes ribosomal protein L16) were further analyzed . Whole chromosomal DNA or PCR products of the 23S ribosomal DNA (rDNA) operons from these mutants could be used to transform the susceptible S . pneumoniae strain R6 to resistance at frequencies of 10(-5) and 10(-4), respectively, rates 10- to 100-fold lower than that for a single-allele chromosomal marker . The transformants appeared slowly (48 to 72 h) on selective medium, and primary transformants passaged on nonselective medium produced single colonies that displayed heterogeneous susceptibilities to evernimicin . A single passage on selective medium of colonies derived from a single primary transformant homogenized the resistance phenotype . Sequence analysis of the 23S rDNA and rRNA from the resistant mutants revealed single, unique mutations in each isolate at the equivalent Escherichia coli positions 2469 (A --> C), 2480 (C --> T), 2535 (G --> A), and 2536 (G --> C) . The mutations map within two different stems of the peptidyltransferase region of domain V . Because multiple copies of rDNA are present in the chromosome, gene conversion between mutant and wild-type 23S rDNA alleles may be necessary for stable resistance . Additionally, none of the characterized mutants showed cross-resistance to any of a spectrum of protein synthesis inhibitors, suggesting that the target site of evernimicin may be unique.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 3049 - 54
Fluoroquinolone resistance in clinical isolates of Streptococcus pneumoniae: contributions of type II topoisomerase mutations and efflux to levels of resistance; Bast DJ et al.; We report on amino acid substitutions in the quinolone resistance-determining region of type II topisomerases and the prevalence of reserpine-inhibited efflux for 70 clinical isolates of S . pneumoniae for which the ciprofloxacin MIC is >/=4 microgram/ml and 28 isolates for which the ciprofloxacin MIC is </=2 microgram/ml . The amino acid substitutions in ParC conferring low-level resistance (MICs, 4 to 8 microgram/ml) included Phe, Tyr, and Ala for Ser-79; Asn, Ala, Gly, Tyr, and Val for Asp-83; Asn for Asp-78; and Pro for Ala-115 . Isolates with intermediate-level (MICs, 16 to 32 microgram/ml) and high-level (MICs, 64 microgram/ml) resistance harbored substitutions of Phe and Tyr for Ser-79 or Asn and Ala for Asp-83 in ParC and an additional substitution in GyrA which included either Glu-85-Lys (Gly) or Ser-81-Phe (Tyr) . Glu-85-Lys was found exclusively in isolates with high-level resistance . Efflux contributed primarily to low-level resistance in isolates with or without an amino acid substitution in ParC . The impact of amino acid substitutions in ParE was minimal, and no substitutions in GyrB were identified.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 3008 - 11
A comparative In vitro surveillance study of gemifloxacin activities against 2,632 recent Streptococcus pneumoniae isolates from across Europe, North America, and South America . The Gemifloxacin Surveillance Study Research Group; Hoban DJ et al.; From 1997 to 1999, 94 study centers in 15 European, 3 North American, and 2 South American countries contributed 2,632 isolates of Streptococcus pneumoniae to an international antimicrobial susceptibility testing study . Only 62.0% of isolates were susceptible to penicillin, while 22.3% were penicillin intermediate and 15.6% were penicillin resistant . Resistance to trimethoprim-sulfamethoxazole (24.4%), azithromycin (26.0%), and clarithromycin (27.1%) was also highly prevalent . For the penicillin-resistant isolates (n = 411), the MICs at which 90% of isolates are inhibited (MIC(90)s) for gemifloxacin, levofloxacin, ofloxacin, clarithromycin, and azithromycin were 0.03, 1, 2, >16, and >64 microgram/ml, respectively . Similarly, for isolates resistant to both azithromycin and clarithromycin (n = 649), gemifloxacin, levofloxacin, ofloxacin, and penicillin MIC(90)s were 0.03, 1, 2, and 4 microgram/ml, respectively . Overall rates of resistance to trovafloxacin (0.3%), levofloxacin (0.3%), grepafloxacin (0.6%), and ofloxacin (0.7%) were low . For ofloxacin-intermediate and -resistant isolates (n = 142), gemifloxacin had the lowest MIC(90) (0.12 microgram/ml) compared to the MIC(90)s of trovafloxacin (0.5 microgram/ml), grepafloxacin (1 microgram/ml), and levofloxacin (2 microgram/ml) . For all S . pneumoniae isolates tested, gemifloxacin MICs were </=0.5 microgram/ml, suggesting that gemifloxacin has the potential to be used as a treatment for pneumococcal infections, including those arising from isolates resistant to beta-lactams and macrolides.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 2999 - 3002
In vitro activity of the new ketolide telithromycin compared with those of macrolides against Streptococcus pyogenes: influences of resistance mechanisms and methodological factors; Bemer-Melchior P et al.; One hundred and seven clinical isolates of Streptococcus pyogenes, 80 susceptible to macrolides and 27 resistant to erythromycin A (MIC >0.5 microgram/ml), were examined . The erythromycin A-lincomycin double-disk test assigned 7 resistant strains to the M-phenotype, 8 to the inducible macrolide, lincosamide, and streptogramin B resistance (iMLS(B)) phenotype, and 12 to the constitutive MLS(B) resistance (cMLS(B)) phenotype . MICs of erythromycin A, clarithromycin, azithromycin, roxithromycin, and clindamycin were determined by a broth microdilution method . MICs of telithromycin were determined by three different methods (broth microdilution, agar dilution, and E-test methods) in an ambient air atmosphere and in a 5 to 6% CO(2) atmosphere . Erythromycin A resistance genes were investigated by PCR in the 27 erythromycin A-resistant isolates . MICs of erythromycin A and clindamycin showed six groups of resistant strains, groups A to F . iMLS(B) strains (A, B, and D groups) are characterized by two distinct patterns of resistance correlated with genotypic results . A- and B-group strains were moderately resistant to 14- and 15-membered ring macrolides and highly susceptible to telithromycin . All A- and B-group isolates harbored erm TR gene, D-group strains, highly resistant to macrolides and intermediately resistant to telithromycin (MICs, 1 to 16 microgram/ml), were all characterized by having the ermB gene . All M-phenotype isolates (C group), resistant to 14- and 15-membered ring macrolides and susceptible to clindamycin and telithromycin, harbored the mefA gene . All cMLS(B) strains (E and F groups) with high level of resistance to macrolides, lincosamide, and telithromycin had the ermB gene . The effect of 5 to 6% CO(2) was remarkable on resistant strains, by increasing MICs of telithromycin from 1 to 6 twofold dilutions against D-E- and F-group isolates.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 2979 - 84
Genetic diversity of the tet(M) gene in tetracycline-resistant clonal lineages of Streptococcus pneumoniae; Doherty N et al.; The aim of the present study was to examine the stability and evolution of tet(M)-mediated resistance to tetracyclines among members of different clonal lineages of Streptococcus pneumoniae . Thirty-two tetracycline-resistant isolates representing three national (Spanish serotype 14, Spanish serotype 15, and Polish serotype 23F) and one international (Spanish serotype 23F) multidrug-resistant epidemic clones were all found to be tet(M) positive and tet(O), tet(K), and tet(L) negative . These isolates all carried the integrase gene, int, which is associated with the Tn1545-Tn916 family of conjugative transposons . High-resolution restriction analysis of tet(M) products identified six alleles, tet(M)1 to tet(M)6: tet(M)1 to tet(M)3 and tet(M)5 in isolates of the Spanish serotype 14 clone, tet(M)4 in both the Spanish serotype 15 and 23F clones, and tet(M)6, the most divergent allele, in the Polish 23F clone . This indicates that tet(M) variation can occur at the inter- and intraclone levels in pneumococci . Two alleles of int were identified, with int1 being found in all isolates apart from members of the international Spanish 23F clone, which carried int2 . Susceptibility to tetracycline, doxycycline, and minocycline was evaluated for all isolates with or without preincubation in the presence of subinhibitory concentrations of tetracyclines . Resistance to tetracyclines was found to be inducible in isolates of all clones; however, the strongest induction was observed in the Spanish serotype 15 and 23F clones carrying tet(M)4 . Tetracycline was found to be the strongest inducer of resistance, and minocycline was found to be the weakest inducer of resistance.

Antimicrob Agents Chemother, 2000 Nov, 44(11), 2962 - 8
Activities of clinafloxacin, gatifloxacin, gemifloxacin, and trovafloxacin against recent clinical isolates of levofloxacin-resistant Streptococcus pneumoniae; Jorgensen JH et al.; The activities of two investigational fluoroquinolones and three fluoroquinolones that are currently marketed were determined for 182 clinical isolates of Streptococcus pneumoniae . The collection included 57 pneumococcal isolates resistant to levofloxacin (MIC >/= 8 microg/ml) recovered from patients in North America and Europe . All isolates were tested with clinafloxacin, gatifloxacin, gemifloxacin, levofloxacin, and trovafloxacin by the National Committee for Clinical Laboratory Standards broth microdilution and disk diffusion susceptibility test methods . Gemifloxacin demonstrated the greatest activity on a per gram basis, followed by clinafloxacin, trovafloxacin, gatifloxacin, and levofloxacin . Scatterplots of the MICs and disk diffusion zone sizes revealed a well-defined separation of levofloxacin-resistant and -susceptible strains when the isolates were tested against clinafloxacin and gatifloxacin . DNA sequence analyses of the quinolone resistance-determining regions of gyrA, gyrB, parC, and parE from 21 of the levofloxacin-resistant strains identified eight different patterns of amino acid changes . Mutations among the four loci had the least effect on the MICs of gemifloxacin and clinafloxacin, while the MICs of gatifloxacin and trovafloxacin increased by up to six doubling dilutions . These data indicate that the newer fluoroquinolones have greater activities than levofloxacin against pneumococci with mutations in the DNA gyrase or topoisomerase IV genes . Depending upon pharmacokinetics and safety, the greater potency of these agents could provide improved clinical efficacy against levofloxacin-resistant pneumococcal strains.

Protein Expr Purif, 2000 Oct, 20(1), 112 - 23
Expression and purification of histidine-tagged proteins from the gram-positive Streptococcus gordonii SPEX system; Myscofski DM et al.; Streptococcus gordonii (S . gordonii) has been used as a gram-positive bacterial expression vector for secreted or surface-anchored recombinant proteins . Fusion of the gram-positive bacterial N-terminal signal sequence to the target protein is all that is required for efficient export . This system is termed SPEX for Surface Protein EXpression and has been used to express proteins for a variety of uses . In this study, the SPEX system has been further developed by the construction of vectors that express polyhistidine-tagged fusion proteins . SPEX vectors were constructed with an N-terminal or C-terminal histidine tag . The C-repeat region (CRR) from Streptococcus pyogenes M6 protein and the Staphylococcus aureus nuclease A (NucA) enzyme were tested for expression . The fusion proteins were purified using metal affinity chromatography (MAC) . Results show that the fusion proteins were expressed and secreted from S . gordonii with the His tag at either the N- or C-terminal position and could be purified using MAC . The M6 fusions retained immunoreactivity after expression and purification as determined by immunoblots and ELISA analyses . In addition, NucA fusions retained functional activity after MAC purification . The M6-His and NucA-His fusions were purified approximately 15- and 10-fold respectively with approximately 30% recovery of protein using MAC . This study shows that the polyhistidine tag in either the N- or C-terminal position is a viable way to purify secreted heterologous proteins from the supernatant of recombinant S . gordonii cultures . This study further illustrates the value of the SPEX system for secreted expression and purification of proteins .

Curr Microbiol, 2000 Mar, 40(3), 149 - 56
Streptococcal opacity factor: a family of bifunctional proteins with lipoproteinase and fibronectin-binding activities; Katerov V et al.; The serum opacity factor (SOF) of Streptococcus pyogenes is a type-specific lipoproteinase of unknown biological significance . We have sequenced the sof gene and characterized the corresponding SOF protein from a strain of type M63 . It was found that sof63 is related to sof22 and that, similar to SOF22 {25}, SOF63 binds fibronectin . Moreover, we demonstrate opacity factor activity in a Streptococcus dysgalactiae fibronectin-binding protein FnBA that is structurally related to the SOF proteins of S . pyogenes . Sequence analysis of these three SOF proteins showed a unique periodical pattern of conserved and variable regions . The enzymatically active part of SOF63 was localized to the fragment corresponding to the entire set of conserved and variable sequences, while for fibronectin-binding a single repeat in the C terminal part of the protein was sufficient . The results show that streptococcal SOF proteins form a novel family of bifunctional proteins with lipoproteinase and fibronectin-binding activities.

Infect Immun, 2000 Nov, 68(11), 6370 - 7
SclA, a novel collagen-like surface protein of Streptococcus pyogenes; Rasmussen M et al.; Surface proteins of Streptococcus pyogenes are important virulence factors . Here we describe a novel collagen-like surface protein, designated SclA (streptococcal collagen-like surface protein) . The sclA gene was identified in silico using the Streptococcal Genome Sequencing Project with the recently identified protein GRAB as the probe . SclA has a signal sequence and a cell wall attachment region containing the prototypic LPXTGX motif . The surface-exposed part of SclA contains a unique NH(2)-terminal domain of 73 amino acids, followed by a collagen-like region . The sclA gene was found to be positively regulated by Mga, a transcriptional activator of several S . pyogenes virulence determinants . A mutant lacking cell wall-associated SclA was constructed and was found to be as effective as wild-type bacteria in platelet aggregation, survival in fresh human blood, and adherence to pharyngeal cells . The sclA gene was found in all 12 S . pyogenes strains that were investigated using PCR . Sequence analysis revealed that the signal sequence and the cell wall attachment region are highly conserved . The collagen-like domain is variable in its NH(2)-terminal region and has conserved repeated domains in its COOH-terminal part . SclA proteins from most strains have additional proline-rich repeats spacing the collagen-like domain and the cell wall attachment sequence . The unique NH(2)-terminal region is hypervariable, but computer predictions indicate a common secondary structure, with two alpha helices connected by a loop region . Immune selection may explain the hypervariability in the NH(2)-terminal region, whereas the preserved secondary structure implies that this region has a common function . These features and the Mga regulation are shared with the M protein of S . pyogenes . Moreover, as with the gene encoding the M protein, phylogenetic analysis indicates that horizontal gene transfer has contributed to the evolution of sclA.

Diagn Microbiol Infect Dis, 2000 Oct, 38(2), 119 - 21
A novel mutation in the alpha-helix 1 of the C subunit of the F(1)/F(0) ATPase responsible for optochin resistance of a Streptococcus pneumoniae clinical isolate; Cogne N et al.; Previously reported mutations involved in optochin resistance of Streptococcus pneumoniae clinical isolates changed residues 48, 49 or 50, in the transmembrane alpha-helix 2 of the F(1)/F(0) ATPase subunit . We report here an unusual mutation which changes the sequence of the transmembrane alpha-helix 1 of the AtpC subunit . This mutation involves a Gly to Ser substitution resulting from a G to A transition at codon 14 of the atpC gene.

Cochrane Database Syst Rev . 2000;(4):CD000023.
Antibiotics for sore throat; Del Mar CB et al.; BACKGROUND: Sore throat is a very common reason for people to attend for medical care . It is a disease that remits spontaneously, that is, 'cure' is not dependent on treatment . Nonetheless primary care doctors commonly prescribe antibiotics for sore throat and other upper respiratory tract infections . OBJECTIVES: To assess the benefits of antibiotics in the management of sore throat . SEARCH STRATEGY: Systematic search of the literature from 1945 to 1999, using electronic searches of MEDLINE (using the keywords, "pharyngitis", "sore throat" and "tonsillitis") after 1966, the Cochrane Library, the Cochrane collection of hand-searched trials, and the reference sections of the articles identified . Abstracts of identified articles were used to determine which studies were trials . SELECTION CRITERIA: Trials of antibiotic against control with either measures of the typical symptoms (throat soreness, headache or fever), or complications (suppurative and non-suppurative) of sore throat . DATA COLLECTION AND ANALYSIS: RevMan 4.0.3 MAIN RESULTS: 25 studies were included in the review . A total number of 11, 452 cases of sore throat have been studied . 1 . Non-suppurative complications There was a trend for protection against acute glomerulonephritis by antibiotics, but insufficient cases were recorded to be sure of this effect . Several studies found benefit from antibiotics for acute rheumatic fever, which reduced this complication to less than one third (OR = 0.30; 95% CI = 0.20-0.45) . 2 . Suppurative complications Antibiotics reduced the incidence of acute otitis media to about one quarter of that in the placebo group (OR = 0.22; 95% CI = 0.11-0.43) and reduced the incidence of acute sinusitis to about one half of that in the placebo group (OR = 0.46; 95% CI = 0.10-2.05) . The incidence of quinsy was also reduced in relation to placebo group (OR = 0.16; 95% CI = 0.07-0.35) . 3 . Symptoms Symptoms of headache, throat soreness and fever were reduced by antibiotics to about one half . The greatest time for this to be evident was at about three and a half days (when the symptoms of about 50% of untreated patients had settled) . About 90% of treated and untreated patients were symptom-free by one week . 4 . Subgroup analyses of symptom reduction Subgroup analysis by age; blind vs unblinded; or use of antipyretics yielded no significant differences . The results of swabs of the throat for Streptococcus influenced the effect of antibiotics . If the swab was positive, antibiotics were more effective (the OR reduced to 0.16, 95% CI 0.09, 0.26) than if it was negative (OR 0.65; 95% CI 0.38,1.1.2) . REVIEWER'S CONCLUSIONS: Antibiotics confer relative benefits in the treatment of sore throat . However, the absolute benefits are modest . Protecting sore throat sufferers against suppurative and non-suppurative complications in modern Western society can only be achieved by treating many with antibiotics who will derive no benefit . Antibiotics shorten the duration of symptoms, but by a mean of only one day about half way through the illness (the time of maximal effect), and by about sixteen hours overall.

J Immunol, 2000 Oct 1, 165(7), 3923 - 33
Group B Streptococcus induces apoptosis in macrophages; Fettucciari K et al.; Group B Streptococcus (GBS) is a pathogen that has developed some strategies to resist host immune defenses . Because phagocytic killing is an important pathogenetic mechanism for bacteria, we investigated whether GBS induces apoptosis in murine macrophages . GBS type III strain COH31 r/s (GBS-III) first causes a defect in cell membrane permeability, then at 24 h, apoptosis . Apoptosis was confirmed by several techniques based on morphological changes and DNA fragmentation . Cytochalasin D does not affect apoptosis, suggesting that GBS-III needs not be within the macrophage cytoplasm to promote apoptosis . Inhibition of host protein synthesis prevents apoptosis, whereas inhibition of caspase-1 or -3, does not . Therefore, GBS can trigger an apoptotic pathway independent of caspase-1 and -3, but dependent on protein synthesis . Inhibition of apoptosis by EGTA and PMA, and enhancement of apoptosis by calphostin C and GF109203X suggests that an increase in the cytosolic calcium level and protein kinase C activity status are important in GBS-induced apoptosis . Neither alteration of plasma membrane permeability nor apoptosis were induced by GBS grown in conditions impeding hemolysin expression or when we used dipalmitoylphosphatidylcholine, which inhibited GBS beta-hemolytic activity, suggesting that GBS beta-hemolysin could be involved in apoptosis . beta-Hemolysin, by causing membrane permeability defects, could allow calcium influx, which initiates macrophage apoptosis . GBS also induces apoptosis in human monocytes but not in tumor lines demonstrating the specificity of its activity . This study suggests that induction of macrophage apoptosis by GBS is a novel strategy to overcome host immune defenses.

FEMS Microbiol Lett, 2000 Sep 15, 190(2), 317 - 21
Neither the A- nor B-repeat regions of the fibrinogen-binding protein of Streptococcus equi subsp . equi are essential for fibrinogen binding; Meehan M et al.; The major cell wall-associated protein (FgBP) of Streptococcus equi subsp . equi possesses two internal blocks of repeated sequence (A and B) and binds horse fibrinogen (Fg) avidly through residues located in the N-terminal half of the molecule . In the present study, we investigated the roles of the two repeats blocks in Fg binding through construction of recombinant FgBP proteins containing defined internal deletions of sequence . Ligand binding experiments clearly showed that neither repeat is essential for Fg binding . However, residues within the B repeats seem to play a major role in the aberrant mobility observed for FgBP following sodium dodecyl sulfate polyacrylamide gel electrophoresis.

FEMS Microbiol Lett, 2000 Sep 15, 190(2), 231 - 6
Allelic variation in a peptide-inducible two-component system of Streptococcus pneumoniae; Reichmann P et al.; The peptide SpiP of Streptococcus pneumoniae regulates the induction of a complex signal transduction system spiR1spiR2spiH . Distinct alleles of spiP and the receptor histidine protein kinase gene spiH were recognized in different pneumococcal clones . The spi system in strain KNR7/87 is adjacent to a bacteriocin gene cluster encoding putative double glycine-type bacteriocins, immunity proteins, and translocator proteins . A direct repeat element upstream of the spiR1 promoter and another three potential transcription start sites within the bacteriocin cluster indicate that SpiP functions as an inducing peptide for bacteriocin synthesis in S . pneumoniae.

CMAJ, 2000 Oct 3, 163(7), 811 - 5
The validity of a sore throat score in family practice; McIsaac WJ et al.; BACKGROUND: Reducing the number of antibiotic prescriptions given for common respiratory infections has been recommended as a way to limit bacterial resistance . This study assessed the validity of a previously published clinical score for the management of infections of the upper respiratory tract accompanied by sore throat . The study also examined the potential impact of this clinical score on the prescribing of antibiotics in community-based family practice . METHODS: A total of 97 family physicians in 49 Ontario communities assessed 621 children and adults with a new infection of the upper respiratory tract accompanied by sore throat and recorded their prescribing decisions . A throat swab was obtained for culture . The sensitivity and specificity of the score approach in this population were compared with previously published results for patients seen at an academic family medicine centre . In addition, physicians' prescribing practices and their recommendations for obtaining throat swabs were compared with score-based recommendations . RESULTS: Of the 621 cases of new upper respiratory tract infection and sore throat, information about prescriptions given was available for only 619; physicians prescribed antibiotics in 173 (27.9%) of these cases . Of the 173 prescriptions, 109 (63.0%) were given to patients with culture-negative results for group A Streptococcus . Using the score to determine management would have reduced prescriptions to culture-negative patients by 63.7% and overall antibiotic prescriptions by 52.3% (both p < 0.01) . Culturing of throat samples would have been reduced by 35.8% (p < 0.01) . There was no statistically significant difference in the sensitivity or specificity of the score approach between this community-based population (sensitivity 85.0%, specificity 92.1%) and an academic family medicine centre (sensitivity 83.1%, specificity 94.3%) . INTERPRETATION: An explicit clinical score approach to the management of patients presenting with an upper respiratory tract infection and sore throat is valid in community-based family practice and could substantially reduce the unnecessary prescribing of antibiotics for these conditions.

J Clin Invest, 2000 Oct, 106(8), 995 - 1002
CD44 as a receptor for colonization of the pharynx by group A Streptococcus; Cywes C et al.; The pharynx is the primary reservoir for strains of group A STREPTOCOCCUS: (GAS) associated both with pharyngitis (streptococcal sore throat) and with invasive or "flesh-eating" soft tissue infections . We now report that CD44, a hyaluronic acid-binding protein that mediates human cell-cell- and cell-extracellular matrix-binding interactions, functions as a receptor for GAS colonization of the pharynx in vivo . We found that attachment of GAS to murine epithelial keratinocytes was mediated by binding of the GAS hyaluronic acid capsular polysaccharide to CD44 . In studies of transgenic mice with a selective defect in epithelial expression of CD44, GAS adherence to CD44-deficient keratinocytes in vitro was reduced compared with adherence to keratinocytes expressing normal levels of CD44 . After intranasal inoculation, GAS colonized the oropharynx of wild-type mice but failed to colonize transgenic mice deficient in CD44 expression . GAS colonization of wild-type mice could be blocked by coadministration of mAb to CD44 or by pretreatment of the animals with exogenous hyaluronic acid . These results provide evidence that CD44 serves as a receptor for GAS colonization of the pharynx and support the potential efficacy of disrupting the interaction between the GAS hyaluronic acid capsule and CD44 as a novel approach to preventing pharyngeal infection.

EMBO J, 2000 Oct 16, 19(20), 5281 - 7
Crystal structure of Streptococcus pneumoniae acyl carrier protein synthase: an essential enzyme in bacterial fatty acid biosynthesis; Chirgadze NY et al.; Acyl carrier protein synthase (AcpS) catalyzes the formation of holo-ACP, which mediates the essential transfer of acyl fatty acid intermediates during the biosynthesis of fatty acids and lipids in the cell . Thus, AcpS plays an important role in bacterial fatty acid and lipid biosynthesis, making it an attractive target for therapeutic intervention . We have determined, for the first time, the crystal structure of the Streptococcus pneumoniae AcpS and AcpS complexed with 3'5'-ADP, a product of AcpS, at 2.0 and 1.9 A resolution, respectively . The crystal structure reveals an alpha/beta fold and shows that AcpS assembles as a tightly packed functional trimer, with a non-crystallographic pseudo-symmetric 3-fold axis, which contains three active sites at the interface between protomers . Only two active sites are occupied by the ligand molecules . Although there is virtually no sequence similarity between the S.pneumoniae AcpS and the Bacillus subtilis Sfp transferase, a striking structural similarity between both enzymes was observed . These data provide a starting point for structure-based drug design efforts towards the identification of AcpS inhibitors with potent antibacterial activity.

J Perinat Med, 2000, 28(4), 309 - 15
Neonatal sepsis of vertical transmission: an epidemiological study from the "Grupo de Hospitales Castrillo"; Lopez Sastre JB et al.; A prospective multicenter study was designed to assess the epidemiology of neonatal sepsis of vertical transmission in Spain . The study was carried out by the "Grupo de Hospitales Castrillo" that included the neonatal services of 19 tertiary care (reference) hospitals and 9 secondary care hospitals . Prospective data from infants with culture-proved neonatal sepsis, clinical sepsis and bacteremia were recorded for 1995 to 1997 . In a total of 203,288 neonates, proven sepsis was diagnosed in 515 (rate of 2.5 per 1000 live births), clinical sepsis in 724 (rate of 3.6 per 1000 live births), and bacteremia of vertical transmission in 155 (rate of 0.76 per 1000 live births) . Very low birth weight (VLBW) infants (< or = 1500 g) showed a significantly higher incidence of confirmed sepsis (26.5 per 1000 live births) and clinical sepsis (32.4 per 1000 live births) than infants weighing > 1500 g . Streptococcus agalactiae was the most frequent causative pathogen in cases of proven sepsis (51%) and bacteremia (33%), but Escherichia coli was the most frequently recovered organism in the VLBW group . The mortality rate of proven sepsis was significantly higher than that of clinical sepsis (8.7% versus 4.3%) (P < 0.01) . In the VLBW cohort, there were no significant differences in the mortality rate between proven sepsis and clinical sepsis . In conclusion, clinical sepsis was the most frequent diagnosis, probably related to intrapartum chemoprophylaxis . Streptococcus agalactiae was the most frequent causative pathogen of culture-positive sepsis and bacteremia, whereas E . coli was the most significant in VLBW infants.

Microb Pathog, 2000 Nov, 29(5), 311 - 7
Mutation in csrR global regulator reduces Streptococcus pyogenes internalization; Jadoun J et al.; Transposon (Tn 916) mutagenesis was employed to identify genes in group A streptococcus (GAS) that are involved in bacterial internalization by epithelial cells . One mutant displayed significantly reduced internalization efficiency and was therefore selected for further characterization . The mutant harbored a single Tn 916 insertion in csr, a genetic locus encoding a two-component regulatory system . Mutations in csr were found to derepress hyaluronic acid (HA) capsule synthesis . Since capsule expression has been previously reported to interfere with internalization of GAS, it was possible that the transposon exerted its inhibitory effect either by derepression of capsule synthesis, or by another mechanism . To study the effect of the csr mutation on bacterial internalization, isogenic mutants deficient in either csrR, hasA or both were generated . The hasA mutant adhered to and internalized into HEp-2 cells significantly better than the parent and the csrR mutant strains . The internalization efficiency of the double mutant (csrR(-)/hasA(-)) was reduced by seven-fold compared to that of the hasA mutant . These findings suggest that csrR affects streptococcal entry by modulating capsule expression as well as by another, yet unknown, mechanism .

J Bacteriol, 2000 Nov, 182(21), 6192 - 202
Gene expression analysis of the Streptococcus pneumoniae competence regulons by use of DNA microarrays; Peterson S et al.; Competence for genetic transformation in Streptococcus pneumoniae is coordinated by the competence-stimulating peptide (CSP), which induces a sudden and transient appearance of competence during exponential growth in vitro . Models of this quorum-sensing mechanism have proposed sequential expression of several regulatory genes followed by induction of target genes encoding DNA-processing-pathway proteins . Although many genes required for transformation are known to be expressed only in response to CSP, the relative timing of their expression has not been established . Overlapping expression patterns for the genes cinA and comD (G . Alloing, B . Martin, C . Granadel, and J . P . Claverys, Mol . Microbiol . 29:75-83, 1998) suggest that at least two distinct regulatory mechanisms may underlie the competence cycle . DNA microarrays were used to estimate mRNA levels for all known competence operons during induction of competence by CSP . The known competence regulatory operons, comAB, comCDE, and comX, exhibited a low or zero initial (uninduced) signal, strongly increased expression during the period between 5 and 12 min after CSP addition, and a decrease nearly to original values by 15 min after initiation of exposure to CSP . The remaining competence genes displayed a similar expression pattern, but with an additional delay of approximately 5 min . In a mutant defective in ComX, which may act as an alternate sigma factor to allow expression of the target competence genes, the same regulatory genes were induced, but the other competence genes were not . Finally, examination of the expression of 60 candidate sites not previously associated with competence identified eight additional loci that could be induced by CSP.

J Bacteriol, 2000 Nov, 182(21), 6154 - 60
Inhibitory effect of heterologous ribosome recycling factor on growth of Escherichia coli; Atarashi K et al.; Ribosome recycling factor (RRF) of Thermotoga maritima was expressed in Escherichia coli from the cloned T . maritima RRF gene and purified . Expression of T . maritima RRF inhibited growth of the E . coli host in a dose-dependent manner, an effect counteracted by the overexpression of E . coli RRF . T . maritima RRF also inhibited the E . coli RRF reaction in vitro . Genes encoding RRFs from Streptococcus pneumoniae and Helicobacter pylori have been cloned, and they also impair growth of E . coli, although the inhibitory effect of these RRFs was less pronounced than that of T . maritima RRF . The amino acid sequence at positions 57 to 62, 74 to 78, 118 to 122, 154 to 160, and 172 to 176 in T . maritima RRF differed totally from that of E . coli RRF . This suggests that these regions are important for the inhibitory effect of heterologous RRF . We further suggest that bending and stretching of the RRF molecule at the hinge between two domains may be critical for RRF activity and therefore responsible for T . maritima RRF inhibition of the E . coli RRF reaction.

J Bacteriol, 2000 Nov, 182(21), 6055 - 65
Defects in D-alanyl-lipoteichoic acid synthesis in Streptococcus mutans results in acid sensitivity; Boyd DA et al.; In the cariogenic organism, Streptococcus mutans, low pH induces an acid tolerance response (ATR) . To identify acid-regulated proteins comprising the ATR, transposon mutagenesis with the thermosensitive plasmid pGh9:ISS1 was used to produce clones that were able to grow at neutral pH, but not in medium at pH 5.0 . Sequence analysis of one mutant (IS1A) indicated that transposition had created a 6.3-kb deletion, one end of which was in dltB of the dlt operon encoding four proteins (DltA-DltD) involved in the synthesis of D-alanyl-lipoteichoic acid . Inactivation of the dltC gene, encoding the D-alanyl carrier protein (Dcp), resulted in the generation of the acid-sensitive mutant, BH97LC . Compared to the wild-type strain, LT11, the mutant exhibited a threefold-longer doubling time and a 33% lower growth yield . In addition, it was unable to initiate growth below pH 6.5 and unadapted cells were unable to survive a 3-h exposure in medium buffered at pH 3.5, while a pH of 3.0 was required to kill the wild type in the same time period . Also, induction of the ATR in BH97LC, as measured by the number of survivors at a pH killing unadapted cells, was 3 to 4 orders of magnitude lower than that exhibited by the wild type . While the LTA of both strains contained a similar average number of glycerolphosphate residues, permeabilized cells of BH97LC did not incorporate D-{(14)C}alanine into this amphiphile . This defect was correlated with the deficiency of Dcp . Chemical analysis of the LTA purified from the mutant confirmed the absence of D-alanine-esters . Electron micrographs showed that BH97LC is characterized by unequal polar caps and is devoid of a fibrous extracellular matrix present on the surface of the wild-type cells . Proton permeability assays revealed that the mutant was more permeable to protons than the wild type . This observation suggests a mechanism for the loss of the characteristic acid tolerance response in S . mutans.

Rev Med Suisse Romande, 2000 Aug, 120(8), 651 - 9
{Pneumococcal antibiotic resistance}; Moreillon P et al.; In 1875, 7 years prior to the description of the Koch bacillus, Klebs visualized the first Streptococcus pneumoniae in a pleural fluid . Since then, this organism has played a determinant role in biomedical science . From a biological point of view, it was largely implicated in the development of passive and active immunization by serotherapy and vaccination, respectively . Genetic transformation was also first observed in S . pneumoniae, leading to the discovery of DNA . From a clinical point of view, S . pneumoniae is still today a prime cause of otitis media in children and of pneumonia in all age groups, as well as a predominant cause of meningitis and bacteremia . In adults, bacteremia is still entailed with a mortality of over 25% . Although S . pneumoniae remained very sensitive to penicillin for many years, penicillin-resistance has emerged and increased dramatically over the last 15 years . During this period of time, the frequency of penicillin-resistant isolates has increased from < or = 1% to frequencies varying from 20 to 60% in geographic areas as diverse as South Africa, Spain, France, Hungary, Iceland, Alaska, and numerous regions of the United States and South America . In Switzerland, the current frequency of penicillin-resistant pneumococci ranges between 5 and > or = 10% . The increase in penicillin-resistant pneumococci correlates with the intensive use of beta-lactam antibiotics . The mechanism of resistance is not due to bacterial production of penicillinase, but to an alteration of the bacterial target of penicillin, the so-called penicillin-binding proteins . Resistance is subdivided into (i) inter mediate level resistance (minimal inhibitory concentration {MIC} of penicillin of 0.1-1 mg/L) and (ii) high level resistance (MCI > or = 2 mg/L) . The clinical significance of intermediate resistance remains poorly defined . On the other hand, highly resistant strains were responsible for numerous therapeutical failures, especially in cases of meningitis . Antibiotics recommended against penicillin-resistant pneumococci include cefotaxime, ceftriaxone, imipenem and in some instances vancomycin . However, penicillin-resistant pneumococci tend to present cross-resistances to all the antibotics of the beta-lactam family and could even become resistant to the last resort drugs mentioned above . Thus, in conclusion, the explosion of resistance to penicillin in pneumococci is a ubiquitous phenomenon which must be fought against by (i) a strict utilization of antibiotics, (ii) the practice of microbiological sampling of infected foci before treatment, (iii) the systematic surveillance of resistance profiles of pneumococci against antibiotics and (iv) the adequate vaccination of populations at risk.

Vaccine, 2000 Oct 15, 19(4-5), 492 - 7
Induction of mucosal and systemic antibody specific for SeMF3 of Streptococcus equi by intranasal vaccination using a sucrose acetate isobutyrate based delivery system; Nally JE et al.; Streptococcus equi causes equine strangles, a highly contagious disease of the upper respiratory tract . The antiphagocytic surface protein SeM is strongly immunogenic and evokes mucosal and systemic antibodies during convalescence . The present study investigated the potential of sucrose acetate isobutyrate (SAIB); a high viscosity excipient that provides controlled release of biologically active substances, to enhance antibody responses following intranasal immunization of horses with a 108 a.a . peptide of SeM (SeMF3) . SeMF3-SAIB was administered intranasally to each of the 11 adult horses on days 0 and 28 . A second group of seven horses was vaccinated with SeMF3 alone . SAIB enhanced the mucosal and systemic immunogenicity of SeMF3, whereas SeMF3 by itself stimulated only a shortlived mucosal IgA and no systemic response . Moreover, nasal mucosal responses of horses immunized with SeMF3-SAIB were qualitatively and quantitatively similar to those observed in convalescent horses and involved similar linear epitopes of SeM . Epitope analysis also suggested that the nasal response was different from that observed in serum . A booster response was obtained after the second vaccination . These results suggest that SAIB has potential as a vehicle for intranasal immunization of horses with antigenic peptides.

J Am Acad Child Adolesc Psychiatry, 2000 Oct, 39(10), 1313 - 5
An open trial of plasma exchange in childhood-onset obsessive-compulsive disorder without poststreptococcal exacerbations; Nicolson R et al.; Patients with childhood-onset obsessive-compulsive disorder (OCD) with symptom exacerbations following streptococcal infections benefit from treatment with plasma exchange . In this study, 5 patients with treatment-refractory OCD without a history of streptococcus-related exacerbations underwent an open 2-week course of therapeutic plasma exchange . Behavioral ratings, completed at baseline and 4 weeks after the initial treatment, included the Clinical Global Impressions Scale and the Yale-Brown Obsessive Compulsive Scale . All 5 patients completed the trial with few side effects, but none showed significant improvement . Plasma exchange does not benefit children and adolescents with OCD who do not have streptococcus-related exacerbations.

Proteins, 2000 Nov 15, 41(3), 323 - 33
Structural consequences of an amino acid deletion in the B1 domain of protein G; O'Neil KT et al.; We describe the NMR structure of a deletion mutant of the B1 IgG-binding domain from Group G Streptococcus . The deletion occurs within the last beta-strand of the protein, where it may potentially have a deleterious effect on the stability of the protein if the protein were not able to conformationally adjust to the perturbation . In particular, the deletion changes the registry of the final three residues in the sheet, forcing a polar Thr to be buried in the interior of the protein and exposing a hydrophobic Val to solvent . The deletion could also potentially create a large cavity in the beta-sheet and force the alpha- and gamma-carboxylates of the C-terminal Glu residue into a partially buried region of the sheet . The structure of the mutant illustrates how the conformation of the protein adjusts to the deletion, thereby mitigating some of the potentially deleterious consequences . Although the elements of secondary structure are retained between the mutant and the wt domain, there are multiple small adjustments in the segments connecting secondary structure elements . In particular, a hydrogen bond between the Glu57 carboxylates and two main chain amides is introduced that alters the conformation in the loop connecting the helix to strand 3 . In addition, to minimize hydrophobic surface exposure, the turn connecting strands 1 and 2 folds toward the core so that the molecular volume is decreased.

J Surg Oncol, 2000 Sep, 75(1), 51 - 4
The influence of enhanced postoperative inflammation by the intrapleural administration of streptococcal preparation (OK-432) on the prognosis of completely resected non-small-cell lung cancer; Uehara T et al.; BACKGROUND AND OBJECTIVES: It is not clear whether postoperative inflammation affects the prognosis of malignant disease . METHODS: We retrospectively reviewed the patients with non-small-cell lung cancer who underwent a complete resection at the National Kyushu Cancer Center from 1989 to 1996 . For the treatment of prolonged air leakage after a pulmonary lobectomy, 25 patients received an intrapleural injection of OK-432, a lyophilized preparation of the heat- and penicillin-treated Su-strain of the Streptococcus pyogenes group A3 . All patients were males who were older than 50 years of age . As a control, we selected 164 male patients who were older than 50 years of age and not given OK-432 during the same period . RESULTS: The administration of OK-432 in most patients was performed on the 4th day after the operation . Pleural drainage could be terminated in a mean of 5.5 days after the intrapleural administration of OK-432 . In the control group, the serum C-reactive protein (CRP) level reached a peak on day 4 after the operation and returned to almost a normal level on day 14 after the operation . In the OK-432 group, the peak CRP level, which was significantly higher than that in the control group, was observed on day 7 after the operation and the elevated CRP level was maintained until 28 days after the operation . The mean level of CRP in the OK-432 group was significantly higher than that in the control on days 7, 14, and 28 after the operation . No significant difference was observed in the disease-free survivals between the two groups . CONCLUSIONS: Based on the above findings, postoperative prolonged inflammation does not seem to affect the progression of subclinically residual tumor cells .

Clin Microbiol Rev, 2000 Oct, 13(4), 588 - 601
Limiting the spread of resistant pneumococci: biological and epidemiologic evidence for the effectiveness of alternative interventions; Schrag SJ et al.; Streptococcus pneumoniae infections are a leading cause of respiratory illness in young children, the elderly, and persons with chronic medical conditions . The emergence of multidrug-resistant pneumococci has compromised the effectiveness of antibiotic therapy for pneumococcal infections . As antibiotic-resistant strains increase in prevalence, there is a need for interventions that minimize the spread of resistant pneumococci . In this review we provide a framework for understanding the spread of pneumococcal resistance and evaluate proposed interventions to reduce this spread . Pneumococci differ from many drug-resistant pathogens because asymptomatic carriers play a key role in transmission of resistant strains and the genes encoding resistance are spread primarily by transformation and conjugative transposons . Evidence suggests that modifications of treatment regimens that have proved effective at limiting resistance in other pathogens may not prevent the spread of pneumococcal resistance . In contrast, programs encouraging more judicious antibiotic use have been shown to be effective . Additionally, a newly developed conjugate pneumococcal vaccine holds great potential as an "antiresistance vaccine" that simultaneously reduces the burden of invasive disease and the prevalence of resistant strains . Several areas of future epidemiologic and laboratory research hold promise to contribute to the reduced spread of pneumococcal resistance.

J Invertebr Pathol, 2000 Oct, 76(3), 222 - 6
Intestinal bacteria affect growth of Bacillus thuringiensis in larvae of the oriental tea tortrix, Homona magnanima diakonoff (Lepidoptera: tortricidae); Takatsuka J et al.; Spores and parasporal crystals of a Bacillus thuringiensis serovar aizawai were fed to fifth instar larvae of the oriental tea tortrix, Homona magnanima, that had been reared aseptically or that had been reared normally . Viable cell numbers of B . thuringiensis and other bacteria in H . magnanima larvae were estimated by homogenization of samples and dilution plating on peptone-polymyxin agar medium for B . thuringiensis cells and on nutrient agar medium for the other bacterial cells . B . thuringiensis did not grow in the larval cadavers of normally reared H . magnanima while bacteria other than B . thuringiensis grew rapidly . In contrast, B . thuringiensis within the larval cadavers of aseptically reared H . magnanima grew and increased 20 times . The bacteria other than B . thuringiensis from the sample homogenates of normally reared larvae that were fed on B . thuringiensis-treated diets had the same characteristics as the bacteria isolated from the guts of healthy H . magnanima larvae, which were putatively identified as Streptococcus spp . and Staphylococcus spp., typical intestinal bacteria of insects . The results strongly suggest that intestinal bacteria influence the growth of B . thuringiensis in the larvae .

J Biol Chem, 2001 Jan 19, 276(3), 2037 - 46 Epub 2000 Oct 06.
Topological organization of the hyaluronan synthase from Streptococcus pyogenes; Heldermon C et al.; Since we first reported (DeAngelis, P . L., Papaconstantinou, J., and Weigel, P . H . (1993) J . Biol . Chem . 268, 19181-19184) the cloning of the hyaluronan (HA) synthase from Streptococcus pyogenes (spHAS), numerous membrane-bound HA synthases have been discovered in both prokaryotes and eukaryotes . The HASs are unique among enzymes studied to date because they mediate 6-7 discrete functions in order to assemble a polysaccharide containing hetero-disaccharide units and simultaneously effect translocation of the growing HA chain through the plasma membrane . To understand how the relatively small spHAS performs these various functions, we investigated the topological organization of the protein utilizing fusion analysis with two reporter enzymes, alkaline phosphatase and beta-galactosidase, as well as several other approaches . From these studies, we conclude that the NH2 terminus and the COOH terminus, as well as the major portion of a large central domain are localized intracellularly . The first two predicted membrane domains were confirmed to be transmembrane domains and give rise to a very small extracellular loop that is inaccessible to proteases . Several regions of the large internal central domain appear to be associated with, but do not traverse, the membrane . Following the central domain, there are two additional transmembrane domains connected by a second small extracellular loop that also is inaccessible to proteases . The COOH-terminal approximately 25% of spHAS also contains a membrane domain that does not traverse the membrane and may contain extensive re-entrant loops or amphipathic helices . Numerous membrane associations of this latter COOH-terminal region and the central domain may be required to create a pore-like structure through which a growing HA chain can be extruded to the cell exterior . Based on the high degree of similarity among Class I HAS family members, these enzymes may have a similar topological organization for their spHAS-related domains.

J Infect Dis, 2000 Nov, 182(5), 1561 - 5 Epub 2000 Oct 09.
Effects of antibiotic class on the macrophage inflammatory response to Streptococcus pneumoniae; Orman KL et al.; Antibiotic choice can alter host inflammation during invasive bacterial infections . Previous studies of gram-negative organisms concluded that antibiotic-mediated release of bacterial cell wall components amplifies inflammation . Less has been reported about antibiotic effect on gram-positive organisms . This study explored the hypothesis that Streptococcus pneumoniae would induce greater macrophage inflammatory mediator production when killed with cell wall active antibiotics rather than protein synthesis inhibitors . Stimulation of RAW 264.7 murine macrophages with pneumococci and oxacillin led to significantly higher inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF) accumulation than did the same concentrations of pneumococci and clindamycin . Neither antibiotic alone or in combination with lipopolysaccharide acted directly on macrophages to modify the immune response . Endotoxin contamination did not confound the results, as preincubation with polymyxin B did not change iNOS or TNF protein levels . Thus, the antimicrobial mechanism of action affects macrophage inflammatory mediator production after stimulation with pneumococci.

J Infect Dis, 2000 Nov, 182(5), 1417 - 24 Epub 2000 Oct 09.
The emergence of Streptococcus pneumoniae resistant to macrolide antimicrobial agents: a 6-year population-based assessment; Gay K et al.; From 1994 through 1999, the available isolates (4148 isolates) from active population-based surveillance of invasive pneumococcal disease in metropolitan Atlanta were serotyped and were tested for antimicrobial susceptibility . Macrolide-resistant isolates were studied for the presence of ermAM (a ribosomal methylase gene), mefE (a macrolide efflux gene), and tetM (the class M tetracycline resistance gene) . Macrolide resistance increased from 16% of all invasive isolates in 1994 to 32% in 1999 . Of the macrolide-resistant pneumococcal isolates studied, 99% contained genomic copies of mefE or ermAM . Isolates with ermAM were mainly serotypes 6B, 23F, 14, or 19F and contained tetM; mefE-associated isolates were predominantly serotypes 14, 6A, or 19F, and most did not contain tetM . The frequency of the ermAM-mediated phenotype in invasive Streptococcus pneumoniae remained stable over the 6-year surveillance . However, the mefE-mediated phenotype increased from 9% in 1994 to 26% of all isolates in 1999 and was noted in new serotypes . By 1999, 93% of the mefE-containing strains had minimum inhibitory concentrations >/=8 microgram/mL . Dissemination of the mefE determinant accounted for the rapid increase in the rate of macrolide resistance in our S . pneumoniae population.

Fam Pract, 2000 Oct, 17(5), 435 - 41
Selections from current literature . Recent advances in conjugated pneumococcal vaccination; Dabelstein D et al.; This article reviews the current literature and recent updates with regard to childhood vaccination for Streptococcus pneumonia including: studies in immunology with antibody titres, dosages of conjugated vaccines, carriage rates of Streptococcus, side effects, comparison in certain disease states and comparison between vaccines.

J Antimicrob Chemother, 2000 Oct, 46(4), 629 - 32
Efficacy of a novel tetracycline derivative, glycylcycline, against penicillin-resistant Streptococcus pneumoniae in a mouse model of pneumonia; Murakami K et al.; The MIC90 of glycylcycline (< or =0.06 mg/L) against 55 strains of Streptococcus pneumoniae was 100-fold lower than that of minocycline or tetracycline . In a mouse model of penicillin-resistant S . pneumoniae (PRSP) pneumonia, glycylcycline (10 mg/kg) decreased bacterial counts in the lungs from 10(6) cfu to <10(2) cfu, whereas no apparent reduction of bacterial numbers was observed with minocycline or penicillin G . Pharmacokinetic studies showed that the half-life and area under the curve of glycylcycline were superior to those of minocycline and penicillin G in the lungs . These results show a preferential distribution of glycylcycline in the lungs and potent in vivo bactericidal activity in PRSP pneumonia.

J Antimicrob Chemother, 2000 Oct, 46(4), 621 - 4
Antimicrobial resistance in Streptococcus pyogenes isolates in Berlin; Arvand M et al.; A total of 212 clinical Streptococcus pyogenes isolates were tested for susceptibility to various antibiotics by agar dilution . The overall frequency of erythromycin resistance was 12.7%, being higher in isolates from children (18.9%) than in those from adult patients (10.7%) . Similar results were found for clarithromycin, while 2.8% of the isolates were resistant to ciprofloxacin . All strains were susceptible to penicillin and cefotaxime . Of the erythromycin-resistant isolates subjected to the double-disc diffusion test for erythromycin and clindamycin, 35% expressed constitutive and 55% inducible resistance to clindamycin . Ten per cent were susceptible to clindamycin (M-phenotype) . Thus, a high rate of macrolide resistance in S . pyogenes has emerged in Berlin.

J Antimicrob Chemother, 2000 Oct, 46(4), 557 - 64
Streptococcus pneumoniae in community-acquired respiratory tract infections in Spain: the impact of serotype and geographical, seasonal and clinical factors on its susceptibility to the most commonly prescribed antibiotics . The Spanish Surveillance Group for Respiratory Pathogens; Marco F et al.; Resistance of Streptococcus pneumoniae to antimicrobial agents shows geographical variation and also depends on serotype, patient age and sample origin . Factors affecting antibiotic resistance in S . pneumoniae were studied from results of a multicentre susceptibility study of 12 antimicrobial agents, carried out in 14 Spanish hospitals between May 1996 and April 1997 . The most frequent serotypes were serotypes 6 (14.8%), 19 (11.8%), 23 (10.5%), 3 (9.9%), 9 (8.0%), 14 (8.0%), 15 (3.9%) and 11 (3.2%); 9.6% of isolates were not typeable . The remaining 25 serotypes constituted 20.2% of strains . Penicillin resistance was more prevalent in serotypes 14 (88%), 23 (66%), 9 (62%) and 6 (57%), whereas erythromycin resistance was more prevalent in serotypes 6 (68.5%), 15 (61.4%), 14 (49.5%), 19 (42.7%) and 23 (39.3%) . Serotypes 6 and 19 were the serotypes most commonly isolated from both children and adults, although with different proportions (24.0% and 19.2% of isolates from children were serotype 6 and 19, respectively, with the corresponding figures for isolates from adults being 13.6% and 11.5%, respectively) . The rates of resistance of pneumococcal strains to penicillin and cefuroxime were significantly higher in strains from children than in those from adults (resistance to penicillin, 50.4% in children and 37.0% in adults; resistance to cefuroxime, 62.4% in children and 45.6% in adults) . There was significantly more resistance to erythromycin in middle ear isolates (48.9%) than in blood isolates (27%) . The prevalence of resistance to beta-lactams showed a seasonal pattern, with higher rates in summer and winter, proportional to the magnitude of resistance . Susceptibility to macrolides did not vary seasonally . Our results stress the relative importance that geographical, temporal (seasonality), patient (sample type, origin, age group) and bacterium-related (serotype) factors have on the variations in susceptibility observed among different pneumococcal clinical isolates.

Jpn J Antibiot, 2000 Jul, 53(7), 512 - 21
{Antimicrobial susceptibility and serotypes of Streptococcus pneumoniae isolated from the patients with respiratory tract infections in Hokusetsu Area of Osaka}; Sugita K et al.; One hundred and thirteen strains of Streptococcus pneumoniae (S . pneumoniae) were isolated from the clinical specimens of patients with respiratory tract infections between January and December 1998 in three hospitals in Hokusetsu area of Osaka . We investigated susceptibility of 113 strains of S . pneumoniae to benzylpenicillin (PCG) and other antimicrobial agents and their serotypes . 1) Of the 113 strains of S . pneumoniae isolated, 25.7% were susceptible (PSSP), 51.3% were intermediate (PISP) and 23% were resistant to benzylpenicillin (PRSP) . 2) The MICs of cefaclor, cefditoren, cefpodoxime, cefdinir, erythromycin, clindamycin and minocycline were elevated, but the MIC values of cefditoren ranged from < or = 0.03 to 1.0 microgram/ml . The susceptibility of 113 strains to cefditoren was comparatively high . 3) The MIC values of imipenem, meropenem and vancomycin for 81 strains of PISP and PRSP ranged from < or = 0.015 to 1.0 microgram/ml, from < or = 0.015 to 2.0 micrograms/ml and from 0.13 to 0.5 microgram/ml, respectively . The susceptibility of these strains to three antimicrobial agents was superior to that to the other antimicrobial agents examined . 4) Of the 60 strains examined, 19, 6, and 23 serotypes were 30, 25 and 18.3%, respectively . The three serotypes were observed in PISP and PRSP with a high frequency . 5) Isolates of S . pneumoniae were 37.2% for children under 2 years of age and 30.9% for children from 2 to 6 years of age . Most of the strains isolated from these children were resistant.

Urology . 2000 Oct 1;56(4):669.
Invasive group A streptococcus infection of the scrotum and streptococcal toxic shock syndrome; Walker BR et al.; We report a case of invasive group A streptococcus infection of the scrotum that presented as epididymoorchitis and rapidly progressed to streptococcal toxic shock syndrome . The presentation, pathophysiology, and management of invasive group A streptococcus and streptococcal toxic shock syndrome are reviewed . Rapid recognition is necessary to avoid the significant morbidity and mortality associated with these invasive infections.

J Microbiol Methods, 2000 Oct, 42(2), 197 - 201
An improved vector system for insertional gene inactivation inspired by the tmRNA-tagging system of S . pneumoniae; Molnos J et al.; Insertional mutagenesis is a technique often used to inactivate genes in Streptococcus pneumoniae . Using conventional vectors, a 5' segment of the targeted gene remains under the control of the gene's authentic promoter following gene disruption . Thus, the expression of a functional peptide and the misinterpretation of results in consequence cannot be excluded . To circumvent this problem, we have developed a plasmid for insertional mutagenesis based on the tmRNA-tagging system of S . pneumoniae which ensures that any protein expressed after gene disruption is degraded . Insertional mutagenesis using this vector results in the targeted gene being tagged with a tmRNA-derived sequence coding for a proteolysis tag . Here we show that the translation product of a gene tagged by this method is not detectable by Western blotting, suggesting that the protein was degraded . This modified vector allows total inactivation of genes with a reliability that cannot be achieved by conventional vectors for insertional mutagenesis . This approach can be applied to other bacterial species.

Clin Chem, 2000 Oct, 46(10), 1583 - 7
Serum procalcitonin concentrations in term delivering mothers and their healthy offspring: a longitudinal study; Assumma M et al.; BACKGROUND: The reported sensitivities and specificities of procalcitonin (PCT) concentrations for the diagnosis of neonatal infection vary widely . A postnatal increase of PCT has been observed in healthy term newborns with a peak at approximately 24 h of age, and many questions remain regarding maternal and perinatal factors that may influence the normal PCT kinetics during the immediate postnatal period . METHODS: We prospectively investigated the association between the serum PCT values obtained from 121 mothers at delivery and serum PCT in their healthy, term offspring at birth as well as at 24 and 48 h of age . We also analyzed whether obstetric and perinatal factors would alter maternal and neonatal PCT response . RESULTS: PCT concentrations in the babies at birth were significantly higher than in the mothers (P <0.0001), with even larger differences at 24 and 48 h of age . None of the variables identified from maternal and perinatal histories had a significant effect on maternal PCT response . In the healthy neonate, the variables that significantly affected the concentration of PCT at birth were the mothers' PCT (P <0.01), maternal group B streptococcus colonization (P <0.05), and rupture of membranes >/=18 h (P <0.01) . The coefficient of linear correlation between the mother's PCT concentration and that of the baby at birth was 0 . 32 (P <0.01) . The only variable that significantly altered the PCT concentration at both 24 (P <0.01) and 48 (P <0.01) h of age was rupture of membranes >/=18 h . Nonetheless, the PCT response observed during the 48-h period after birth among healthy babies born to mothers with risk factors for infection was well below that reported previously among age-matched neonates with sepsis . CONCLUSIONS: The postnatal increase of PCT observed in the healthy neonate with peak values at 24 h of age most likely represents endogenous synthesis . In estimating the sensitivities and specificities of PCT for diagnosis of sepsis throughout the initial 48 h of life, it is important to consider the normal PCT kinetics and the pattern(s) of PCT response in the healthy neonate.

Nat Biotechnol, 2000 Oct, 18(10), 1060 - 4
Therapy of mucosal candidiasis by expression of an anti-idiotype in human commensal bacteria; Beninati C et al.; Two recombinant strains of Streptococcus gordonii, secreting or displaying a microbicidal single-chain antibody (H6), and stably colonizing rat vagina, were used to treat an experimental vaginitis caused by Candida albicans . A post-challenge intravaginal delivery of the H6-secreting strain was as efficacious as fluconazole in rapidly abating the fungal burden . Three weeks after challenge, 75% and 37.5% of the rats treated with the H6-secreting or displaying bacteria, respectively, were cured of the infection, which persisted in 100% of the animals treated with a S . gordonii strain expressing an irrelevant single-chain antibody . Thus, a human commensal bacterium can be suitably engineered to locally release a therapeutic antibody fragment.

Pediatrics, 2000 Oct, 106(4), 695 - 9
Pneumococcal mastoiditis in children; Kaplan SL et al.; OBJECTIVE: To determine the impact of antibiotic resistance on the frequency, clinical features, and management/outcome of mastoiditis attributable to Streptococcus pneumoniae . DESIGN: Retrospective review of the medical records of children with mastoiditis caused by S pneumoniae from September 1993 through December 1998 . PATIENTS: Infants and children with pneumococcal mastoiditis cared for at 8 children's hospitals in the United States . RESULTS: Thirty-four children with pneumococcal mastoiditis were identified . The median age of the children was 12 months (range: 2 months-12.5 years); 28 (82%) were </=2 years old . Six children had recurrent otitis media . A subperiosteal abscess was noted in 13 children (37%) . The mastoids were abnormal in all 25 patients on whom computed tomography was performed . There was no trend toward increasing numbers of cases per year despite increasing proportions of pneumococcal isolates, which were nonsusceptible to penicillin . Serogroup 19 accounted for 57% of isolates, serogroup 23 for 14.3% of isolates, and serotype 3 for 10 . 7% of isolates . Except for receipt of less antibiotic therapy in the previous 30 days, children with penicillin-susceptible isolates had similar demographic features and clinical findings and surgical treatment as did children whose isolates were nonsusceptible to penicillin . CONCLUSIONS: Pneumococcal mastoiditis occurs primarily in children <2 years of age and usually is not associated with a history of recurrent otitis media . The number of cases of mastoiditis caused by S pneumoniae occurring among 8 children's hospitals has remained stable despite increasing rates of antibiotic-resistant S pneumoniae . Serogroup 19 is the leading serogroup associated with pneumococcal mastoiditis.Streptococcus pneumoniae, mastoiditis, serotypes, resistance.

J Infect Dis, 2000 Nov, 182(5), 1425 - 36 Epub 2000 Oct 03.
Human immune response to streptococcal inhibitor of complement, a serotype M1 group A Streptococcus extracellular protein involved in epidemics; Hoe NP et al.; Streptococcal inhibitor of complement (Sic) is a highly polymorphic extracellular protein made by serotype M1 group A Streptococcus strains that contributes to bacterial persistence in the mammalian upper respiratory tract . New variants of the Sic protein arise very rapidly by positive selection in human populations during M1 epidemics . The human antibody response to Sic was analyzed . Of 636 persons living in diverse localities, 43% had anti-Sic serum antibodies, but only 16.4% had anti-M1 protein serum antibody . Anti-Sic antibody was also present in nasal wash specimens in high frequency . Linear B cell epitope mapping showed that serum antibodies recognized epitopes located in structurally variable regions of Sic and the amino terminal hypervariable region of the M1 protein . Phage display analyses confirmed that the polymorphic regions of Sic are primary targets of host antibodies . These results support the hypothesis that selection of Sic variants occurs on mucosal surfaces by a mechanism that involves acquired host antibody.

Caries Res, 2000 Sep-Oct, 34(5), 427 - 31
Binding characteristics of Streptococcus mutans for calcium and casein phosphopeptide; Rose RK; Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP) and may be used to localize ACP in dental plaque, maintaining a state of supersaturation with respect to tooth enamel, reducing demineralization and enhancing remineralization {Reynolds, J Dent Res 1997;76:1587-1595} . The aim of this paper is to investigate these effects by measuring the affinity and capacity of Streptococcus mutans for CPP-ACP . Using the equilibrium dialysis system described by Rose and Hogg {Biochim Biophys Acta 1995;1245:94-98}, assessment of calcium binding by a plaque streptococcus at a fixed CPP-ACP concentration gives a series of CPP-ACP-influenced dissociation constants for calcium . These data can then be used to derive a true dissociation constant for CPP-ACP itself . The results demonstrate that CPP-ACP binds with about twice the affinity of the bacterial cells for calcium up to a value of 0 . 16 g/g wet weight cells . Application of CPP-ACP to plaque may cause a transient rise in plaque fluid free calcium which may assist remineralization . Subsequently, CPP-ACP will form a source of readily available calcium to inhibit demineralization . Hence, CPP-ACP binds well to plaque, providing a large calcium reservoir, which is likely to restrict mineral loss during a cariogenic episode and provide a potential source of calcium for subsequent remineralization . Overall, once in place, CPP-ACP will restrict the caries process.

Caries Res, 2000 Sep-Oct, 34(5), 418 - 26
Effects of Apis mellifera propolis on the activities of streptococcal glucosyltransferases in solution and adsorbed onto saliva-coated hydroxyapatite; Koo H et al.; Propolis, a resinous hive product collected by Apis mellifera bees, has been used for thousands of years in folk medicine . Ethanolic extracts of propolis (EEP) have been shown to inhibit the activity of a mixture of crude glucosyltransferase (Gtf) enzymes in solution . These enzymes synthesize glucans from sucrose, which are important for the formation of pathogenic dental plaque . In the present study, the effects of propolis from two different regions of Brazil on the activity of separate, purified Gtf enzymes in solution and on the surface of saliva-coated hydroxyapatite (sHA) beads were evaluated . The EEP from Minas Gerais (MG; Southeastern Brazil) and Rio Grande do Sul (RS; Southern Brazil) were tested for their ability to inhibit the enzymes GtfB (synthesis of insoluble glucan), GtfC (insoluble/soluble glucan) and GtfD (soluble glucan) . The effects of propolis on Gtf from Streptococcus sanguis (soluble glucan synthesis) was also explored . The EEP from both regions effectively inhibited the activity of all Gtfs in solution (75-95%) and on the surface of sHA beads (45-95%) at concentrations between 0.75 and 3.0 mg of propolis/ml . However, the two samples of propolis showed different levels of inhibition on each of the enzymes tested . In general, EEP RS demonstrated a significantly higher inhibitory activity on GtfB and C activities (both solution and surface assays) than EEP MG at concentrations between 0.047 and 0.187 mg/ml (p<0.05) . EEP MG, on the other hand, exhibited a greater inhibitory effect on the activities of surface GtfD (at 0.375, 0.75 and 1.5 mg/ml) and S . sanguis Gtf (at 1.5 and 3.0 mg/ml; p<0.05) . These data indicate that EEP is a potent inhibitor of Gtf enzymes in solution and adsorbed on an experimental pellicle; however, its effect on Gtf activity is variable depending on the geographical origin of the propolis samples . There is a need to identify the active compounds of propolis.

Caries Res, 2000 Sep-Oct, 34(5), 412 - 7
Differences in acidogenicity of S . sobrinus and S . rattus are linked to the catalytic efficiency of the glycolytic key enzyme phosphofructokinase; Gansser G et al.; This contribution describes the biochemical properties of two catalytically different phosphofructokinases (PFKs) purified from Streptococcus rattus LB 2 (PFK-rat) and Streptococcus sobrinus OMZ 65 (PFK-sob), respectively . Steady-state kinetics revealed K(M) = 0 . 8 mM for PFK-rat and K(M) = 0.08 mM for PFK-sob for F-6-P as the substrate . The enzymes also differ in their pH profiles: whereas the highest activity of PFK-rat was measured at pH = 8.0, the optimum pH of PFK-sob was at pH = 7.0 . In addition, compared to PFK-sob, PFK-rat was more sensitive against the allosteric inhibitor ATP . PFK catalyzes a committed step of glycolysis, the main acid producing catabolic pathway . Thus, the catalytically more efficient enzyme isolated from S . sobrinus OMZ 65, especially at low pH, could explain the comparably high acidogenicity of this strain.

Curr Microbiol, 2000 Nov, 41(5), 336 - 40
Effects of thymol on ruminal microorganisms; Evans JD et al.; Thymol (5-methyl-2-isopropylphenol) is a phenolic compound that is used to inhibit oral bacteria . Because little is known regarding the effects of this compound on ruminal microorganisms, the objective of this study was to determine the effects of thymol on growth and lactate production by the ruminal bacteria Streptococcus bovis JB1 and Selenomonas ruminantium HD4 . In addition, the effect of thymol on the in vitro fermentation of glucose by mixed ruminal microorganisms was investigated . Neither 45 nor 90 microg/ml of thymol had any significant effect on growth or lactate production by S . bovis JB1, but 180 microg/ml of thymol completely inhibited growth and lactate production . In the case of S . ruminantium HD4, 45 microg/ml of thymol had little effect on growth and lactate production; however, 90 microg/ml of thymol completely inhibited growth of S . ruminantium HD4 . Thymol also decreased glucose uptake by whole cells of both bacteria . When mixed ruminal microorganisms were incubated in medium that contained glucose, 400 microg/ml of thymol increased final pH and the acetate to propionate ratio and decreased concentrations of methane, acetate, propionate, and lactate . In conclusion, thymol was a potent inhibitor of glucose fermentation by S . bovis JB1 and S . ruminantium HD4 . Even though thymol treatment decreased methane and lactate concentrations and increased final pH in mixed ruminal microorganism fermentations of glucose, concentrations of acetate and propionate were also reduced.

Eur J Clin Microbiol Infect Dis, 2000 Aug, 19(8), 627 - 30
Molecular typing of bacteria directly from cerebrospinal fluid; Enright MC et al.; Using Streptococcus pneumoniae as an example, the ability of multilocus sequence typing (MLST) to characterise isolates directly from cerebrospinal fluid (CSF) was investigated . A nested multiplex polymerase chain reaction method that amplifies the seven housekeeping gene fragments used for pneumococcal MLST was applied to 30 CSF samples from suspected cases of bacterial meningitis . The fragments were amplified from all 14 samples from which Streptococcus pneumoniae was cultured, and, after direct sequencing, the allelic profiles obtained from ten of the samples corresponded to those of clones previously associated with invasive pneumococcal disease . MLST could also predict the penicillin susceptibility and serotype of the CSF isolates.

Eur J Clin Microbiol Infect Dis, 2000 Aug, 19(8), 618 - 22
Evaluation of the Vitek 2 system for susceptibility testing of Streptococcus pneumoniae isolates; Goessens WH et al.; Vitek 2 (bioMerieux, France) is a new commercial system that allows rapid identification and rapid determination of the minimum inhibitory concentration (MIC) of Streptococcus pneumoniae by monitoring the growth kinetics of the organisms in microwells . The accuracy of the Vitek 2 system in susceptibility testing was evaluated by determining the MICs of 50 penicillin-susceptible and 150 intermediate or penicillin-resistant Streptococcus pneumoniae isolates and comparing the results with those obtained using the agar dilution method . The essential agreement between the Vitek 2 system and the reference method was 91% for penicillin, 93% for cefotaxime and ceftriaxone, and more than 94% for amoxicillin, erythromycin, ofloxacin, co-trimoxazole, tetracycline, and imipenem . One very major error (1.1%) and one major error (0.9%) were obtained for tetracycline . The minor error rate for penicillin of 19.3% was mainly due to intermediate category isolates (n = 29) being identified as resistant and susceptible isolates (n = 6) being identified as intermediate by the commercial system . The minor error rates for amoxicillin, cefotaxime, ceftriaxone, imipenem, and ofloxacin were 25.4%, 25.4%, 29.4%, 19.2%, and 31.5%, respectively . Vancomycin, tetracycline, co-trimoxazole, and erythromycin showed minor error rates of 0-6.1% . In conclusion, Vitek 2 shows good agreement with the reference method, as demonstrated by the low numbers of major errors, but it has a tendency to overestimate MICs, resulting in minor errors.

J Int Med Res, 2000 Jul-Aug, 28(4), 187 - 90
Predominant Staphylococcus aureus isolated from various skin diseases; Higaki S et al.; We examined predominant Staphylococcus aureus isolated from lesions of various skin diseases, and during the past 3 years, 113 methicillin-sensitive and 31 methicillin-resistant S . aureus have been isolated . The predominant species isolated from almost all of the primary bacteriological cultures was S . aureus . The skin diseases from which cultures were most frequently prepared were atopic dermatitis, followed by ulcers . S . aureus was the predominant species in two-thirds or more of cases of all of the different skin diseases examined . The predominant species identified, other than S . aureus, included Streptococcus species and Pseudomonas aeruginosa . Evaluation of the predominance of S . aureus is important to the determination of the severity of skin lesions and, if needed, appropriate antimicrobial therapy.

Scand J Immunol, 2000 Oct, 52(4), 348 - 55
Protective effect of Plantago major L . Pectin polysaccharide against systemic Streptococcus pneumoniae infection in mice; Hetland G et al.; The antibacterial effect of a soluble pectin polysaccharide, PMII, isolated from the leaves of Plantago major, was examined in inbred NIH/OlaHsd and Fox Chase SCID mice experimentally infected with Streptococcus pneumoniae serotype 6B . Serotype 6B is known to give a more protracted infection when injected intraperitoneally into susceptible mice than more virulent serotypes like type 4 . PMII was administered i.p . either once 3 days before challenge or once to thrice from 3 to 48 h after challenge . The number of bacteria in blood and the mouse survival rate were recorded . Pre-challenge administration of PMII and also lipopolysaccharide (LPS), included as a control, gave a dose-dependent protective effect against S . pneumoniae type 6B infection . However, injection of PMII after establishment of the infection in NIH/OlaHsd mice had no effect . The data demonstrate that, firstly, the polysaccharide fraction PMII from P . major protects against pneumococcal infection in mice when administered systemically prechallenge, and secondly that the protective effect is owing to stimulation of the innate and not the adaptive immune system.

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