Streptococcal

J Infect Dis, 2004 Mar 1, 189(5), 775 - 84 Epub 2004 Feb 16.
Does the presence of pneumococcal DNA in middle-ear fluid indicate pneumococcal etiology in acute otitis media?
Palmu AA, Saukkoriipi PA, Lahdenkari MI, Kuisma LK, Makela PH, Kilpi TM, Leinonen M.
Bacterial culture of middle-ear fluid (MEF), the standard for etiologic diagnosis of acute otitis media (AOM), has revealed Streptococcus pneumoniae (Pnc) to be a major pathogen responsible for one-third of AOM cases . In the present study, we compared the results of polymerase chain reaction (PCR) based on the amplification of the pneumolysin gene with the results of pneumococcal culture, for 2595 MEF samples obtained during AOM events in 831 children who were followed from 2-24 months of age in the Finnish Otitis Media Vaccine Trial . PCR results were positive for 47.1% of the MEF samples, and culture results were positive for 27.3% of the samples . PCR-positive, culture-negative samples were associated with previous Pnc AOM in a time-dependent pattern, concurrent antibiotic treatment, low volume of MEF, and concurrent nasopharyngeal carriage . PCR-positive AOM represented a clinically less severe disease, compared with culture-positive Pnc AOM . A positive PCR result seemed to indicate the presence of viable, although often nonculturable, Pnc.

Rev Med Chir Soc Med Nat Iasi, 2002 Oct-Dec, 106(4), 796 - 800
{Preclinical assay on a magnetic carrier type ferrofluid}; Bredetean O et al.; The aim of the study was to determine the acute toxicity and the antimicrobial actions of an original magnetic carrier, type ferrofluid . The hydrophilic ferrofluid was prepared by covering the Fe3O4 nannoparticles with ammoniumoleate . The absolute amount of iron was of 40 mg/ml ferrofluid . METHODS: Acute toxicity was evaluated on five groups of Swiss male mice, after a single intraperitoneal administration of 1, 0.75, 0.5, 0.25 and 0.125 ml dose of pure ferrofluid/100 g body weight (b.w.), using step-level toxicity method . The study groups of mice were follow-up for 10 days . We did not use the same volume of solution for all the study groups because we were concerned about not to modify the behavior of the ferrofluid (but for each group we used the same volume of solution) . The tasks of this part of the study were: 1) the record of the mice death in the first 10 days after intraperitoneal administration of ferrofluid; 2) the behavior of the animal subjects; 3) the morphopathologic examination of kidney, lung, heart, liver and peritoneum samples from the death mice and from the after ten days survivors which were sacrificed . We also investigated the possible antibacterial actions of the ferrofluid on E . coli spp., Klebsiella spp., Staphylococcus aureus Streptococcus group D., in the second part of the study, using standard lab kit . The validation of the results was performed using controls for E . colli and Staphylococcus aureus . RESULTS: The death of the mice was registered between 24 and finished after 96 hours . The maximum tolerated dose (MTD), was of 0.25 ml (10 mg iron/100 g b.w.) and the lethal dose hundred percent (LD100) was of 0.75 ml/(30 mg iron/100 b.w.) . In our study we did not determined any kind of antibacterial action of the ferrofluid . CONCLUSIONS: 1) LD100, in our study, was of 30 mg iron/100 g b.w., and DMT of 10 mg iron/100 g b.w . 2) The death of the mice may be due to toxic aggression of ammonium ions released, in vivo, from the ammoniumoleate coverage of magnetite nannoparticles . 3) There were no in vitro antibacterial actions for this ferrofluid.

J Antimicrob Chemother, 2004 Apr, 53(4), 620 - 5 Epub 2004 Feb 18.
Spectrum of antibiotic resistance of the Spain14-5 Streptococcus pneumoniae clone over a 22 year period; Perez-Trallero E et al.; OBJECTIVE: To study the characteristics and the evolution through time of a single Streptococcus pneumoniae multidrug-resistant international clone . METHODS: From 1981 to 2002, the presence of the multidrug-resistant Spain14-5 clone was studied among the 4201 S . pneumoniae isolated in Gipuzkoa (northern Spain) . RESULTS: Overall, 93 isolates belonging to the Spain14-5 clone were identified . The first isolate of this clone was detected in 1981 and was already resistant to beta-lactams, erythromycin, clindamycin and chloramphenicol . The reference strain from the international collection for this clone was susceptible to macrolides and lincosamides whereas most of the isolates studied, including the first isolate detected in 1981, were resistant to macrolides and had the erm(B) gene encoding macrolide resistance . CONCLUSIONS: The clone was genetically stable through time, was multiresistant since its inception and has recently become highly resistant to fluoroquinolones . The characteristic antibiotic resistance pattern of this clone should include erythromycin resistance.

J Bacteriol, 2004 Mar, 186(5), 1398 - 408
Relevance of peptide uptake systems to the physiology and virulence of Streptococcus agalactiae; Samen U et al.; Streptococcus agalactiae is a major cause of invasive infections in human newborns . To satisfy its growth requirements, S . agalactiae takes up 9 of the 20 proteinogenic amino acids from the environment . Defined S . agalactiae mutants in one or several of four putative peptide permease systems were constructed and tested for peptide uptake, growth in various media, and expression of virulence traits . Oligopeptide uptake by S . agalactiae was shown to be mediated by the ABC transporter OppA1-F, which possesses two substrate-binding proteins (OppA1 and OppA2) with overlapping substrate specificities . Dipeptides were found to be taken up in parallel by the oligopeptide permease OppA1-F, by the dipeptide ABC transporter DppA-E, and by the dipeptide symporter DpsA . Reverse transcription-PCR analysis revealed a polycistronic organization of the genes oppA1-F and dppA-E and a monocistronic organization of dpsA in S . agalactiae . The results of quantitative real-time PCR revealed a medium-dependent expression of the operons dppA-E and oppA1-F in S . agalactiae . Growth of S . agalactiae in human amniotic fluid was shown to require an intact dpsA gene, indicating an important role of DpsA during the infection of the amniotic cavity by S . agalactiae . Deletion of the oppB gene reduced the adherence of S . agalactiae to epithelial cells by 26%, impaired its adherence to fibrinogen and fibronectin by 42 and 33%, respectively, and caused a 35% reduction in expression of the fbsA gene, which encodes a fibrinogen-binding protein in S . agalactiae . These data indicate that the oligopeptide permease is involved in modulating virulence traits and virulence gene expression in S . agalactiae.

Arch Pharm Res, 2004 Jan, 27(1), 57 - 60
Anti-allergic activity of 18beta-glycyrrhetinic acid-3-O-beta-D-glucuronide; Park HY et al.; Glycyrrhizin (18beta-glycyrrhetinic acid-3-O-beta-D-glucuronopyranosyl-(1 --> 2)-beta-D-glucuronide, GL) was transformed to 18beta-glycyrrhetinic acid-3-O-beta-D-glucuronide (GAMG) by Streptococcus LJ-22 . The antiallergic activities of GL and GAMG was measured using a RBL cell assay system and contact hypersensitivity model mice . GAMG exhibited anti-allergic activity with IC50 values of 0.28 mM . GAMG, which is sweeter than GL, and 18beta-glycyrrhetinic acid, which is a GAMG metabolite by human intestinal bacteria, also inhibited the passive cutaneous anaphylaxis and skin contact inflammation . In conclusion, GAMG may be useful as a new sweet food additive and an anti-allergic agent.

Ann Ig, 2003 Sep-Oct, 15(5), 567 - 73
{Risks and benefits of influenza and pneumococcal immunization in HIV-1 infected individuals}; Tanzi E et al.; Influenza and Streptococcus pneumoniae diseases can cause severe complications in HIV-1 infected individuals leading to increases in hospital admission and even death . Both vaccinations are recommended for such individuals, but some studies reported that such immunizations may stimulate an increase of HIV-1 viral load and decrease of CD4+ cells count . A review of published studies, including our studies carried out in HIV-1 infected former drugs addicts, indicates that influenza and pneumococcal vaccinations are well tolerated in individuals with HIV-1, and do not induce deterioration of the course of HIV-1 infection, even though the immune response to vaccination is lower than that one observed in immunocompetent individuals . Therefore the lack of significant changes of virological and immunological parameters indicates that such immunizations can be safely administrated to HIV-1 infected individuals.

Biomedica, 2003 Dec, 23(4), 456 - 61
{Effectiveness of the antibiotics chloramphenicol and rifampin in the treatment of Streptococcus pneumoniae-induced meningitis and systemic infections}; Hernandez M et al.; Streptococcus pneumoniae, a common pathogen in pediatric infections, has become resistant to penicillin and make these infections difficult to treat . Rifampin and chloramphenicol have been recommended as alternative therapies, since they are less costly and more accessible to communities with limited resources . However, their use may be restricted by the differing levels of resistance found in target populations . The objective was to determine minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) for chloramphenicol and rifampin in strains of S . pneumoniae . These strains were newly isolated from children under age 5 that had demonstrated systemic infections and meningitis . A subgroup of 107 isolates of S . pneumoniae was selected from 324 strains isolated during a period of 2 years (1994-1996) . Among these isolates, 60 were penicillin-resistant and 47 were susceptible; 53 isolates were from children with meningitis . MIC and MBC for chloramphenicol and rifampicin were obtained by standard methods recommended by the National Committee for Clinical Laboratory Standards (NCCLS) . S . pneumoniae ATCC strain 49619 served as the control . An isolate was considered susceptible to chloramphenicol when MIC = 4 microg/ml and resistant when MIC = 8 microg/ml . A strain was considered susceptible to rifampin when MIC = 1 microg/ml and resistant when MIC = 4 microg/ml . MBC was determined by recording the lower concentration of the antibiotic that inhibited 99.9% of the initial inoculum . Chloramphenicol resistance was found in 21% of the 107 isolates . In the group susceptible to penicillin, 11% were resistant to chloramphenicol and in the group resistant to penicillin 28% was resistant to chloramphenicol as well . MBC was found > 4 microg/ml in 28% of the isolates susceptible to penicillin and in 60% of the resistant isolates . No isolates were found resistant to rifampin . However, 2 penicillin resistant isolates showed CBM > 1 microg/ml to rifampin, and one with CIM = 1 microg/ml had a MBC to rifampicin of 16 microg/ml . Meningitis isolates showed higher CIM and CBM than the group of total isolates . These data suggest that chloramphenicol is not recommended for invasive infections caused by S . pneumoniae in Colombia . Rifampin is a more effective therapy in combination with other antibiotics for treatment of this kind of infections . Further studies are necessary to clarify the significance of low levels of MBC to rifampin found in some strains, since this may affect the efficacy of therapies that include this antibiotic.

Arch Otolaryngol Head Neck Surg, 2004 Feb, 130(2), 201 - 7
Age-, site-, and time-specific differences in pediatric deep neck abscesses; Coticchia JM et al.; OBJECTIVE: To clarify presentations, organisms, and locations of deep neck abscesses in children . DESIGN: Retrospective review . SETTING: Tertiary children's hospital . The study population comprised 169 patients younger than 19 years who were surgically treated for deep neck abscesses between 1989 and 1999 . MAIN OUTCOME MEASURE: Resolution of abscess . RESULTS: Neck mass (91%), fever (86%), cervical adenopathy (83%), poor oral intake (66%), and neck stiffness (59%) were common in all ages . Patients younger than 4 years, compared with patients 4 years or older, presented with agitation (50% vs 14%), cough (35% vs 14%), drooling (27% vs 12%), lethargy (46% vs 33%), oropharyngeal abnormalities (45% vs 60%), respiratory distress (5% vs 2%), retractions (5% vs 2%), rhinorrhea (53% vs 15%), stridor (4% vs 2%), and trismus (14% vs 53%) . Children younger than 1 year were infected with Staphylococcus aureus (79%) vs group A streptococcus (6%) . Children 1 year or older were infected with group A streptococcus (29%) vs S aureus (16%) . Retropharyngeal or parapharyngeal regions were involved in children 1 year or older (49%) vs younger than 1 year (21%) . Anterior or posterior triangles and submandibular or submental regions were involved in 39% and 36%, respectively, of children younger than 1 year vs 30% and 23%, respectively, of children 1 year or older . Retropharyngeal and parapharyngeal abscesses yielded group A streptococcus (34%) vs S aureus (11%) . Anterior and posterior triangle abscesses yielded S aureus (35%) vs group A streptococcus (19%), as did submandibular and submental abscesses (42% vs 19%) . CONCLUSIONS: Abscesses in children younger than 1 year affected anterior or posterior triangles and submandibular or submental regions, yielding S aureus . Abscesses in children 1 year or older affected retropharyngeal or parapharyngeal regions, yielding group A streptococcus.

J Microbiol Methods, 2004 Mar, 56(3), 401 - 12
Expression of green fluorescent protein and its application in pathogenesis studies of serotype 2 Streptococcus suis; Lun S et al.; We investigated the interaction between type 2 Streptococcus suis and swine phagocytes during infection of the natural host, by using green fluorescent protein (GFP) as a specific marker to observe the challenge organism . We compared the strength of the S . suis sly promoter (SP332) and the synthetic promoter (CP25) in driving GFP expression . Two GFP alleles, gfpP11 and gfpmut3*, were also compared . The two promoters and two alleles were efficiently compared using three different promoter-GFP gene combinations on a shuttle vector, which were transformed into S . suis strains SX332, SX932 or M2 . Plasmid pSL6.81 has SP332 with gfpP11, pSL5.24 has SP332 with gfpmut3*, and pSL5.28 has CP25 with gfpmut3* . The transformants were fluorescent with green light when viewed with an epifluorescence microscope or during flow cytometry . The signal was also detected using a laser scanning confocal microscope . The GFP expression level varied and CP25 with gfpmut3* led to greatest expression . For optimizing GFP detection, fluorescence-based cell sorting was applied to SX332(pSL5.28) and the mean fluorescence intensity increased 25.9% after optimization . Fluorescence activated cell sorting (FACS)-based phagocytosis assay showed that, without opsonization, phagocytosis rates of SX332, SX932 and M2 by both neutrophils and monocytes were similar and low . After opsonization, the phagocytosis of M2 increased 10-fold while phagocytosis of SX332 and SX932 did not change . GFP-labeled S . suis was identified in fresh pig tonsil tissue 18 h after infection . The results of this study indicated that GFP was expressed in type 2 S . suis and GFP labeled S . suis could be used in phagocytosis and pathogenesis studies.

J Korean Med Sci, 2004 Feb, 19(1), 21 - 6
Therapeutic efficacy of meropenem for treatment of experimental penicillin-resistant pneumococcal meningitis; Kim SW et al.; With the widespread emergence of antimicrobial resistance, combination regimens of ceftriaxone and vancomycin (C+V) or ceftriaxone and rifampin (C+R) are recommended for empirical treatment of pneumococcal meningitis . To evaluate the therapeutic efficacy of meropenem (M), we compared various treatment regimens in a rabbit model of meningitis caused by penicillin-resistant Streptococcus pneumoniae (PRSP) . Therapeutic efficacy was also evaluated by the final bacterial concentration in the cerebrospinal fluid (CSF) at 24 hr . Each group consisted of six rabbits . C+V cleared the CSF at 10 hr, but regrowth was noted in 3 rabbits at 24 hr . Meropenem monotherapy resulted in sterilization at 10 hr, but regrowth was observed in all 6 rabbits at 24 hr . M+V also resulted in sterilization at 10 hr, but regrowth was observed in 2 rabbits at 24 hr . M+V was superior to the meropenem monotherapy at 24 hr (reduction of 4.8 vs . 1.8 log10 cfu/mL, respectively; p=0.003) . The therapeutic efficacy of M+V was comparable to that of C+V (reduction of 4.8 vs . 4.0 log10 cfu/mL, respectively; p=0.054) . The meropenem monotherapy may not be a suitable choice for PRSP meningitis, while combination of meropenem and vancomycin could be a possible alternative in the treatment of PRSP meningitis.

Di Yi Jun Yi Da Xue Xue Bao, 2004 Feb, 24(2), 168 - 71, 176
Calcium signaling events in Streptococcus pneumoniae invasion of human type II pneumocytes; Xu BL et al.; OBJECTIVE: To study whether Streptococcus pneumoniae (S.pn) can provoke filamentous actin (F-actin) rearrangements in vitro through calcium signaling pathways in type II pneumocytes(A549 cells), resulting in S.pn invasion of the cells . METHOD: After FITC-phalloidin labeling of F-actin, F-actin rearrangements were observed by S.pn adhesion to type pneumocyte A549 cells . S.pn invasion of A549 cells was determined by pretreating A549 cells with cytochalasin D . To investigate whether F-actin rearrangements could be blocked by Ca2+ inhibitors, A549 cells were pretreated with Ca2+ inhibitors dantrolene, and loaded in Fura-2/AM probe to determine the concentration of cytosolic free calcium by S.pn adhesion to A549 cells after 30, 60, and 90 min respectively . RESULTS: Intact S.pn can promote F-actin rearrangements . Cytochalasin D was able to prevent S.pn invasion of A549 cells . No invasion of A549 cell can be determined at 0.25 microg/ml of cytochalasin D . One subset of the inhibitors of Ca2+ signal transduction molecules blocked F-actin rearrangements dose-dependently, and S.pn adhesion of A549 cells for 30, 60, and 90 min increased cytosolic free calcium, reaching 487.5+/-38.1 , 548.2+/-35.6 and 557.2+/-47.5 nmol/L, respectively . They were higher than of the control group . CONCLUSION: S.pn can provoke F-actin rearrangements through Ca2+ signaling pathways, which further leads to S.pn invasion of A549 cells.

Protein Expr Purif, 2003 Dec, 32(2), 232 - 8
Stable isotope labeling of a Group A Streptococcus virulence factor using a chemically defined growth medium; Vise PD et al.; A secreted, hypervariable virulence factor called the streptococcal inhibitor of complement (Sic) has been linked to the reemergence of epidemics due to the human pathogenic bacterium Group A Streptococcus . This paper describes a method for expressing and purifying Sic from an attenuated GAS strain using a chemically defined growth medium . This method was used to label specific amino acid residue types in Sic with forms containing the magnetically active isotope (15)N, at the amide nitrogen . The (15)N-labeling of Sic permits a detailed investigation of the structure and dynamics of the protein using nuclear magnetic resonance spectroscopy . The level of stable isotope incorporation was established using mass spectrometry and nuclear magnetic resonance spectroscopy.

Acta Paediatr Taiwan, 2003 Sep-Oct, 44(5), 274 - 8
Epidemiological and clinical features of group A Streptococcus pharyngitis in children; Lin MH et al.; In order to understand the prevalence of childhood streptococcal pharyngitis, isolation of group A Streptococcus (GAS) was attempted from throat swabs of pharyngitis patients . Children aged between 1 and 15 years presenting to the outpatient department with pharyngeal erythema were prospectively enrolled in the study . Demographic data and presenting symptoms and signs for each patient were recorded and a throat swab was taken . Of 1175 throat cultures obtained, GAS was isolated in 252 cases (21.4%) . Of these, 142 (56.3%) were boys and 110 (43.7%) girls . A higher proportion of boys was found with GAS pharyngitis (1.29: 1) . The mean age of GAS culture-positive patients was 7.8 +/- 2.3 years old . Patients aged between 6 and 11 years were more prevalent in GAS pharyngitis . Ninety (35.7%) of our GAS pharyngitis patients occurred between March and May . A second smaller peak occurred between October and December . The following factors showed independent positive correlation with GAS infection: sore throat (p < 0.001), no coryza (p = 0.011), tonsillar swelling (p < 0.001), anterior cervical adenopathy (p = 0.029) and scarlatiniform rash (p < 0.001) . However, GAS was found in less than half of the patients who had these clinical manifestations . In conclusion, pharyngeal infection with GAS in children is not uncommon . The prevalence of GAS pharyngitis is related to patient gender, age, and month of the year . Diagnosis of GAS pharyngitis based on clinical features alone is unreliable.

New Microbiol, 2004 Jan, 27(1), 83 - 6
Inhibitory effect of water-soluble chitosan on growth of Streptococcus mutans; Fujiwara M et al.; The present study was undertaken to evaluate the effects of pH and the degree of polymerization of chitosan on the inhibition of growth of Streptococcus mutans . Three types of chitosan, polymer, oligomer and monomer, were used at 4% (W/V) and three different levels of pH: 6.0, 6.5 and 7.4 . Bactericidal activity was calculated by the growth ratio . Chitosan oligomer significantly inhibited bacterial growth at a pH value of 6.5 . All three types of chitosan strongly inhibited bacterial growth at pH 6.0 . Furthermore, nearly complete inhibition was obtained with 2%(W/V) chitosan solution at constant pH 6.5 . This study is the first to report that water-soluble chitosan directly suppresses the growth of the typical cariogenic bacterium S . mutans even at pH 6.5, without causing demineralization of the tooth surface.

J Antimicrob Chemother, 2004 Mar, 53(3), 457 - 63 Epub 2004 Feb 12.
Macrolide resistance and genotypic characterization of Streptococcus pneumoniae in Asian countries: a study of the Asian Network for Surveillance of Resistant Pathogens (ANSORP); Song JH et al.; OBJECTIVES: To characterize mechanisms of macrolide resistance among Streptococcus pneumoniae from 10 Asian countries during 1998-2001 . METHODS: Phenotypic and genotypic characterization of the isolates and their resistance mechanisms . RESULTS: Of 555 isolates studied, 216 (38.9%) were susceptible, 10 (1.8%) were intermediate and 329 (59.3%) were resistant to erythromycin . Vietnam had the highest prevalence of erythromycin resistance (88.3%), followed by Taiwan (87.2%), Korea (85.1%), Hong Kong (76.5%) and China (75.6%) . Ribosomal methylation encoded by erm(B) was the most common mechanism of erythromycin resistance in China, Taiwan, Sri Lanka and Korea . In Hong Kong, Singapore, Thailand and Malaysia, efflux encoded by mef(A) was the more common in erythromycin-resistant isolates . In most Asian countries except Hong Kong, Malaysia and Singapore, erm(B) was found in >50% of pneumococcal isolates either alone or in combination with mef(A) . The level of erythromycin resistance among pneumococcal isolates in most Asian countries except Thailand and India was very high with MIC(90)s of >128 mg/L . Molecular epidemiological studies suggest the horizontal transfer of the erm(B) gene and clonal dissemination of resistant strains in the Asian region . CONCLUSION: Data confirm that macrolide resistance in pneumococci is a serious problem in many Asian countries.

J Antimicrob Chemother, 2004 Mar, 53(3), 487 - 93 Epub 2004 Feb 12.
Oral anti-pneumococcal activity and pharmacokinetic profiling of a novel peptide deformylase inhibitor; Gross M et al.; OBJECTIVE: BB-81384, a novel peptide deformylase (PDF) inhibitor, was characterized in terms of enzyme inhibition profile, antibacterial activity, rodent pharmacokinetics and oral efficacy in murine infection models . METHODS: MICs were determined by standard NCCLS broth microdilution . Selectivity of metalloenzyme inhibition was determined with a limited panel of enzymes via standard biochemical assays . Profiling of the pharmacokinetics and select tissue disposition in mice was determined and compared with that of the macrolide, azithromycin . In vivo murine efficacy studies using Streptococcus pneumoniae were conducted using a peritonitis model, as well as lung and thigh burden models of infection . RESULTS: BB-81384 selectively inhibited PDF with an IC(50) approximately 10 nM and with MICs < 0.5 mg/L against most S . pneumoniae pathogens . Pharmacokinetic analysis revealed good oral bioavailability and moderate clearance and volume of distribution . BB-81384 partitioning to lung tissue was similar in terms of magnitude and kinetics to that of the plasma compartment . Single-administration oral efficacy in a mouse peritonitis model was evident with an ED(50) of 30 mg/kg . BB-81384 reduced the bacterial load by approximately 5 and 3 log units in organ-burden models of lung and thigh infection, respectively . CONCLUSION: BB-81384, a novel PDF inhibitor with good activity against S . pneumoniae in vitro, was the first compound of this class to be profiled for oral pharmacokinetics and tissue disposition and to demonstrate oral anti-pneumococcal efficacy in mice.

Am J Ophthalmol, 2004 Feb, 137(2), 308 - 12
An in vivo study comparing the ocular absorption of levofloxacin and ciprofloxacin prior to phacoemulsification; Bucci FA Jr; PURPOSE: To compare aqueous humor concentrations of levofloxacin vs ciprofloxacin when used as prophylactic medications before phacoemulsification . DESIGN: Patients (n = 93) were randomly assigned to receive either 0.5% levofloxacin (Quixin) or 0.3% ciprofloxacin (Ciloxan) using one of the following dosing regimens: (A) 1 to 2 drops four times a day for 2 days preoperatively; (B) 5 doses (1 to 2 drops) delivered every 10 minutes in the hour immediately preceding surgery; or (C) the combination of A and B . METHODS: Aqueous samples (0.1 ml) were obtained immediately before surgery, and drug concentrations were measured using high performance liquid chromatography/mass spectrometry . RESULTS: The mean concentration of levofloxacin in the aqueous humor was significantly greater than that of ciprofloxacin in all treatment groups (P <.001): 284.8 vs 67.4 microg/ml (regimen A); 1,135.6 vs 185.6 microg/ml (regimen B); and 1,618.6 vs 241.5 (regimen C) . Dosing regimen B delivered significantly more drug to the aqueous humor than regimen A for both levofloxacin (P < or =.001) and ciprofloxacin (P =.004) . Dosing regimen C delivered significantly more drug to the aqueous humor than regimen B for levofloxacin (P =.05) but not for ciprofloxacin (P =.384) . CONCLUSIONS: Although the concentration of active drug in levofloxacin is approximately 1.7-fold higher than that in ciprofloxacin, the aqueous concentration of levofloxacin after topical administration was four to seven times greater than ciprofloxacin; these differences were statistically significant . With dosing regimens B and C, levofloxacin concentrations in the aqueous humor were above the MIC90 for most common ocular pathogens, including Staphylococcus and Streptococcus species . Ciprofloxacin did not reach such concentrations in any treatment group.

BJOG, 2004 Mar, 111(3), 226 - 30
Antenatal screening and intrapartum management of Group B Streptococcus in the UK; Kenyon S et al.; OBJECTIVE: To determine whether there has been any change in UK policy for the screening and intrapartum management of Group B Streptococcus in pregnancy over a two year period . DESIGN: Two national survey's of practice carried out in 1999 and 2001 . SETTING: All obstetric units in the UK . POPULATION: Clinical directors of maternity services . METHODS: A questionnaire was sent to all clinical directors of maternity services in the UK requesting information about their policy and practice with respect to antenatal screening for Group B Streptococcus colonisation . Reminders were sent after one month . MAIN OUTCOME MEASURES: Number of maternity units in the UK screening and offering intrapartum antibiotic prophylaxis for Group B Streptococcus colonisation in pregnancy . RESULTS: The response rates were 84% in 1999 and 82% in 2001 . Of the responding units, six (3%) in 1999 and four (2%) in 2001 used vaginal swab based screening for Group B Streptococcus colonisation in the antenatal period . In 1999, intrapartum antibiotic prophylaxis was offered to women with a previous baby affected by Group B Streptococcus in 85% (176/207) of maternity units and in 2001 this had risen to 95% (193/203) . Similarly, in 1999 intrapartum antibiotic prophylaxis was offered to women who were known carriers of Group B Streptococcus in 87% (179/207) of maternity units and in 2001 this had risen to 95% (193/203) . Appropriate dosage of a recommended antibiotic was prescribed in 7% (9/123) units in 1999 and in 20% (35/178) units in 2001 . CONCLUSIONS: Although intrapartum antibiotic prophylaxis for women at high risk of giving birth to babies with Group B Streptococcus is widely practiced in the UK, a programme of antenatal screening for Group B Streptococcus colonisation has not been adopted along the lines advocated in the USA . There therefore remains an opportunity to evaluate such a screening programme in a randomised trial.

Echocardiography, 2004 Feb, 21(2), 171 - 3
Double-chambered right ventricle associated with mural and pulmonic valve endocarditis: description of a clinical case and review of the literature; Lopez-Pardo F et al.; A double-chambered right ventricle is a relatively uncommon congenital cardiac defect characterized by the presence of anomalous muscle bundles dividing the right ventricle into a high-pressure proximal chamber and a low-pressure distal chamber . This pathology is often wrongly diagnosed in adult patients . We report the first case of a patient with double-chambered right ventricle associated with a mural and pulmonic valve endocarditis caused by Streptococcus parasanguis diagnosed with two-dimensional echocardiography . During the course of treatment, the patient suffered from a septic pulmonary embolism, and subsequently required surgical intervention, which confirmed the echocardiographic findings.

Rev Esp Quimioter, 2003 Dec, 16(4), 412 - 20
{Serotypes and antibiotic susceptibility of Streptococcus pneumoniae in children in the health district of Valencia and Castellón, Spain: a multicenter, prospective trial}; Canton E et al.; A study was conducted to determine the serotypes, the coverage of the heptavalent conjugate vaccine (VCN 7-v) and the antibiotic susceptibility of Streptococcus pneumoniae in children (<15 years) in the health districts of the provinces of Castellon and Valencia, Spain, from June 1999 to December 2002 . A total of 271 strains were evaluated, 15.5% of which were invasive, 63.5% were of respiratory origin and 22.5% were from conjunctival and otitic exudates; 67.2% of the strains were found in children younger than 2 years of age . The distribution of the serotypes showed slight changes according to age and the origin of the sample: the most common serotypes were 19, 6, 23, 14, 3, 9 and 11; however, in children younger than 2 years of age the order changed to 19, 6, 14 and 23, 9, 11 and 3, and in the invasive strains to 19, 6, 14, 3 and 23 . A total of 27.2% of the stains were susceptible to the ten antibiotics tested . For penicillin, reduced susceptibility was found in 50.4% and high resistance in 5.8%; 98.3% were susceptible to ampicillin and amoxicillin-clavulanic acid, and 80.7% to cefuroxime; 52.5% were resistant to erythromycin and 43.7% to clindamycin . No strains were resistant to rifampicin, vancomycin, levofloxacin or cefotaxime . The most susceptible serotypes were 3 and 11, and the most resistant was 14 (which consistently showed resistance to an antibiotic), 19, 6 and 23 . According to these data, the theoretical coverage of VCN 7-v would be 80.1% in children younger than 2 years and 73.43% in those aged 0-14 years.

Pediatr Infect Dis J, 2004 Feb, 23(2), 177 - 9
Streptococcus pneumoniae empyema necessitatis; Freeman AF et al.; Empyema necessitatis is a rare complication of empyema characterized by extension of suppuration from the pleural space through the chest wall . The most common etiologies are tuberculosis and actinomycosis . We describe a 1-year-old boy with Streptococcus pneumoniae empyema necessitatis and review previously reported cases.

Pediatr Infect Dis J, 2004 Feb, 23(2), 91 - 8
Tolerability and immunogenicity of an eleven valent mixed carrier Streptococcus pneumoniae capsular polysaccharide-diphtheria toxoid or tetanus protein conjugate vaccine in Finnish and Israeli infants; Dagan R et al.; BACKGROUND: To have wide global coverage of pneumococcal serotypes, the number of serotypes covered by the current 7-valent pneumococcal conjugate vaccine must be increased . We have studied the safety and immunogenicity of an 11-valent mixed carrier vaccine (PncDT11) in infants . METHODS: The study vaccine contained polysaccharide antigens of serotypes 1, 4, 5, 7F, 9V, 19F and 23F conjugated to tetanus protein and serotypes 3, 6B, 14 and 18C conjugated to diphtheria toxoid . The vaccine was administered to Finnish (n = 117) and Israeli (n = 135) infants at ages 2, 4, 6 and 12 months concomitantly with other vaccines used in national vaccination programs . IgG antibodies to polysaccharides were determined by enzyme immunoassay from serum samples taken at ages 2, 7, 12 and 13 months . After each injection the infants were followed for 30 min to detect any immediate adverse reactions, and parents were given a diary card to report any adverse events during the next 5 days . RESULTS: No severe adverse reactions occurred, and immediate adverse reactions were rare . After each dose approximately 30% of the vaccinees experienced local reactions of which pain was the most common . Fever of >38 degrees C was reported in 33 to 53% of the vaccinees and high fever (>40 degrees C) was reported 6 times . The PncDT11 vaccine was immunogenic . The antibody concentrations after primary immunization series were higher in Israeli than in Finnish infants, but the differences were not significant for most serotypes . The difference was most marked at 13 months, a time point at which the difference was significant in 10 of 11 serotypes . CONCLUSION: PncDT11 is safe and immunogenic in infants . The use of 11-valent pneumococcal vaccine would increase the serotype coverage beyond the currently available 7-valent vaccine.

Neurology, 2004 Feb 10, 62(3), 509 - 11
Axonal injury, a neglected cause of CNS damage in bacterial meningitis; Nau R et al.; The contribution of axonal injury to CNS damage in bacterial meningitis was studied by histology and immunohistochemistry for amyloid-beta precursor protein in humans and experimental rabbits . Axonal injury in the white matter caused predominantly but not exclusively by ischemia was detected in all autopsy cases (n = 5) and in 11 of 15 brains of rabbits 18 to 24 hours after intracisternal infection with Streptococcus pneumoniae . This suggests a substantial contribution of axonal pathology to neurologic sequelae after bacterial meningitis.

Oral Microbiol Immunol, 2004 Apr, 19(2), 132 - 5
Distribution and characterization of hemolytic activity by an oral anaerobe from the Streptococcus milleri group; Yamaguchi T et al.; Some oral anaerobes from the Streptococcus milleri strain group were found to secrete human specific hemolytic toxin, which was detected when bacteria were cultured in Todd-Hewitt broth and Brain Heart Infusion broth . The toxin elicited by the Streptococcus intermedius strain was partially fractionated by ammonium sulfate precipitation . Preincubation with glutathione or cysteine showed significant inhibiting effects; however, no effects were seen with dithiothreitol or beta-mercaptoethanol, and cholesterol was a weak inhibitor . Five kinds of protease inhibitor had no effect on the hemolytic activity, and rabbit preimmune and immune sera against the bacterial cells showed weak inhibition at a similar level . Digestion with trypsin, chymotrypsin, proteinase-K, subtilisin and pronase-P brought about a rise in activity, followed by a decrease during long-term incubation . Other enzymes tested showed no effects . Further, the presence of the intermedilysin gene in the portion with hemolytic activity was not identified by polymerase chain reaction.

Oral Microbiol Immunol, 2004 Apr, 19(2), 102 - 5
Roles of Streptococcus mutans dextranase anchored to the cell wall by sortase; Igarashi T et al.; In order to clarify the role that sortase (SrtA) plays in anchoring dextranase (Dex) to the cell wall of Streptococcus mutans, both Dex- and SrtA- mutants were constructed by insertional inactivation of the respective genes . Western blot analysis with a Dex antiserum showed that in the srtA mutant the Dex was not bound to the cell wall but was secreted into the culture supernatant . In contrast, in the wild type, Dex remained cell-wall-associated . Biological properties of the srtA mutant were examined in dextran fermentation, colony morphology and adherence to a smooth surface . The srtA mutant, as well as the wild type, retained the ability to ferment dextran . However, the colony morphology of the srtA mutant on Todd Hewitt agar containing sucrose was much larger than that of the wild type and showed a ring-like structure . In addition, the srtA mutant was more adhesive to a smooth surface than the wild type when sucrose was present . However, the adhesion of the srtA mutant remarkably decreased by addition of exogenous dextranase . These studies suggest that the SrtA mediates Dex-anchoring to the cell wall in S . mutans, and cell wall-anchored Dex plays a role in controlling both the adhesive properties of extracellular glucan and the ability to utilize extracellular glucan as a nutrient source . In contrast, extracellular Dex is only responsible for degrading extracellular glucan as a nutrient source.

Pediatr Infect Dis J, 2004 Feb, 23(2 Suppl), S125 - 8
Pneumococcal resistance in perspective: how well are we combating it?
Amsden GW.
Because Streptococcus pneumoniae is the most commonly isolated community-acquired respiratory tract pathogen, the reports of high rates of antibiotic resistance throughout the world highlight the need for intervention to stem any further increases in resistance . Efforts to reduce the incidence of pneumococcal resistance have been mainly 2-fold, involving attempts to reduce unnecessary antibiotic prescribing, as well as to assure early childhood immunization with the pneumococcal heptavalent conjugate vaccine . To reduce unnecessary prescribing for infections that are typically viral in etiology, such as acute bronchitis, education efforts have been focused not only on clinicians but also on parents and patients . These education efforts significantly reduce unnecessary antibiotic prescribing, and initial evidence suggests that they may stabilize, if not reduce, the incidence of penicillin and macrolide-resistant pneumococcal isolates . Utilization of the relatively new pneumococcal heptavalent conjugate vaccine not only reduces the incidence of acute otitis media caused by pneumococcal serotypes included in the vaccine as well as disease caused by related serotypes but also has a highly significant effect on reducing the incidence of invasive pneumococcal disease in children and potential adult contacts . In addition more recent data have established that vaccination is also decreasing the carriage and transmission of antibiotic-resistant pneumococcal isolates . Education and vaccine programs that attempt to stabilize and/or reduce the rate of pneumococcal resistance are at least as important as having effective antibiotic treatments for pneumococcal disease . These efforts to address pneumococcal resistance have been highly successful to date.

Chest, 2004 Feb, 125(2), 566 - 71
Treatment of complicated parapneumonic pleural effusion with intrapleural streptokinase in children; Yao CT et al.; OBJECTIVE: To evaluate the role of intrapleural streptokinase in the management of complicated parapneumonic effusions in children . DESIGN: Prospective comparative study . SETTING: Cheng Kung University Hospital, a tertiary medical center in Tainan, Taiwan . PATIENTS AND METHODS: We enrolled as our prospective study group 20 consecutive children with complicated parapneumonic effusions who received intrapleural streptokinase treatment between August 2000 and July 2002 . We also retrospectively analyzed a comparison group of 22 consecutive children with complicated parapneumonic effusions who received chest tube drainage without streptokinase treatment from January 1992 to July 2000 . We then compared the clinical manifestations and outcome of these two patient groups . The patient population (21 boys and 21 girls) ranged in age from 9 to 130 months (mean age, 41.5 +/- 26.3 months {mean +/- SD}) . The characters of pleural effusion showed no difference between the two groups . Nineteen patients had positive findings for Streptococcus pneumoniae, 2 patients had positive findings for Staphylococcus aureus, 2 patients had positive findings for Pseudomonas aeruginosa, and 19 patients had undetermined pathogens . All patients were treated with appropriate antibiotics . RESULTS: More pleural fluid was drained from the streptokinase group than from the comparison group during streptokinase treatment (816 +/- 481 mL vs 279 +/- 238 mL, p < 0.01) . The duration of fever after chest tube insertion was also significantly lower in the study group (5.3 +/- 3.1 days vs 7.9 +/- 4.6 days, p < 0.05) . Only two patients in the streptokinase group required surgical intervention compared with nine patients in the comparison group (p < 0.05) . No major side effects were noticed after streptokinase instillation . CONCLUSION: Intrapleural fibrinolytic treatment with streptokinase is safe and effective, and it can obviate the need for surgery in most cases . The combination treatment should be attempted early on, when complicated parapneumonic effusion is first diagnosed.

FEMS Microbiol Lett, 2004 Feb 9, 231(1), 33 - 8
The Mig protein of Streptococcus dysgalactiae inhibits bacterial internalization into bovine mammary gland epithelial cells; Song XM et al.; The role of the Mig protein of Streptococcus dysgalactiae in bacterial adhesion and internalization of bovine mammary gland epithelial cells (MAC-T) was investigated with the wild-type and isogenic mig mutant strains . While there was no difference in adhesion between the strains, the wild-type strain exhibited a significantly lower level of invasion than the mutants . The lower level of internalization of the Mig(+) strain is likely due to Mig-mediated interference with uptake of the microorganisms rather than the host protein binding properties of Mig . Avoidance of intimate interactions with the host cells might be an alternative strategy for S . dysgalactiae to survive and persist in the bovine mammary glands.

FEMS Microbiol Lett, 2004 Feb 9, 231(1), 27 - 32
Effect of pH on the activity of bovicin HC5, a bacteriocin from Streptococcus bovis HC5; Houlihan AJ et al.; The bacteriocin, bovicin HC5, catalyzed potassium efflux from Streptococcus bovis JB1, and this activity was highly pH dependent . When the pH was near neutral, glucose-energized cells were not affected by bovicin HC5, but the intracellular steady-state concentration of potassium decreased at acidic pH values . The idea that pH was affecting bovicin HC5 binding was supported by the observation that acidic pH also enhanced the efflux of potassium from non-energized cells that had been loaded with potassium . The relationship between bovicin HC5 concentration and potassium depletion was a saturation function, but cooperativity plots indicated that the binding of one bovicin molecule to the cell membrane facilitated the binding of another.

Nihon Kokyuki Gakkai Zasshi, 2004 Jan, 42(1), 62 - 7
{Polymicrobial infections in patients with Legionella pneumonia}; Takayanagi N et al.; We assessed the frequency and clinical significance of polymicrobial infections in 31 patients with sporadic community-acquired Legionella pneumonia . Twenty-six patients were men, 5 were women and mean age was 61 years . Eighteen patients were smokers, 6 patients were chronic alcoholics and 23 had underlying diseases . Regarding severity, the illnesses were mild (two patients), moderate (seven patients) and severe (twenty-two patients) . In 9 (29%) of the patients, one other etiologic agent for community-acquired pneumonia was identified in addition to the Legionella species . The distribution of one other causal agent was as follows: Mycoplasma pneumoniae, 2 patients; Chlamydia pneumoniae, 2; Chlamydia psittaci, 1; Influenza virus, 1; Streptococcus pneumoniae, 1; Klebsiella pneumoniae, 1; Pseudomonas aeruginosa, 1 patient . Because an antimicrobial agent with activity against Legionella species can also provide coverage for Mycoplasma pneumoniae . Chlamydia pneumoniae, and Chlamydia psittaci, the patients with these coinfections improved without any complications . The patient with influenzavirus coinfection became seriously ill, and the condition was complicated by disseminated intravascular coagulation, renal failure and aspergillus bronchitis . The case of Pseudomonas aeruginosa coinfection was accompanied with a lung abscess and empyema . Our experience illustrates the importance of considering polymicrobial infections in patients with sporadic community-acquired Legionella pneumonia.

J Basic Microbiol, 2004, 44(1), 66 - 74
Construction of a GBS-GAS DNA subtraction library allows discovery of previously unidentified GBS genes and rapid location of unique regions on the GBS chromosome; Suvorov AN et al.; A subtraction library of group B streptococcus (GBS) strain O9OR with GAS chromosomal DNA (strain SF370) was constructed and more than 100 plasmid clones sequenced . DNA sequences of the plasmid inserts were analyzed using the BLAST gene search . Most inserts had little or no homology to GAS chromosomal DNA and 26 clones from the library had no gene homologues in the gene bank . The majority of genes discovered represented house keeping GBS genes, but several could be considered as possible virulence factors . Inserts from 21 clones were labeled and used as probes for hybridization with GBS DNA fragments separated by pulsed field electrophoresis . A genetic map of GBS strain O9OR was constructed.

J Infect Dis, 2004 Feb 15, 189(4), 711 - 6 Epub 2004 Jan 30.
Improved host defense against pneumococcal pneumonia in platelet-activating factor receptor-deficient mice; Rijneveld AW et al.; Platelet-activating factor (PAF) is a phospholipid with proinflammatory properties that binds to a specific receptor (PAF receptor {PAFR}) that is expressed on many different cell types . PAFR is able to bind phosphorylcholine, which is present in both PAF and the pneumococcal cell wall . Activation of respiratory epithelial cells in vitro results in up-regulation of PAFR, which, in turn, facilitates invasion of Streptococcus pneumoniae . To determine the role of PAFR in host defense against pneumococcal pneumonia, PAFR-deficient (PAFR(-/-)) and wild-type (wt) mice were inoculated intranasally with S . pneumoniae . PAFR(-/-) mice were relatively resistant to pneumococcal pneumonia, as indicated by delayed and reduced mortality, diminished outgrowth of pneumococci in lungs, and reduced dissemination of the infection (all P<.05, vs . wt mice) . PAFR(-/-) mice also had less pulmonary inflammation . These data provide evidence that PAFR is used by S . pneumoniae to induce lethal pneumonia.

Caries Res, 2004 Mar-Apr, 38(2), 79 - 84
Identification of chromosomes associated with dental caries susceptibility using quantitative trait locus analysis in mice; Nariyama M et al.; Dental caries is a multifactorial, infectious disease with little known about the host genetic factors influencing susceptibility . This study aimed to identify the major candidate chromosomes for dental caries susceptibility and to detect the relevant regions within these . Quantitative trait locus (QTL) analysis was performed on genetic crosses of C3H/HeJ (caries-resistant) and C57BL/6J (caries-susceptible) mice inoculated with Streptococcus mutans serotype C . In a genomewide scan, three suggestive QTLs were detected on chromosomes 1, 2, and 7, one significant QTL was found on chromosome 2, and one highly significant QTL was detected on chromosome 8 . The likelihood ratio statistic (LRS) was raised around the marker D1Mit21 in the middle region of chromosome 1, between D2Mit255 and D2Mit311 in the distal region of chromosome 2, and the region distal to D7Mit31 on chromosome 7 . A significant QTL was located between the markers D2Mit237 and D2Mit101 on chromosome 2 . The LRS was highly significantly raised between markers D8Mit208 and D8Mit280 on chromosome 8, and exceeded a highly significant level between markers D8Mit211 and D8Mit280 . These results suggest that major gene(s) responsible for dental caries susceptibility or resistance are located in one or more of these regions .

Microbiology, 2004 Feb, 150(Pt 2), 365 - 71
Mouse skin passage of Streptococcus pyogenes results in increased streptokinase expression and activity; Rezcallah MS et al.; The plasminogen activator streptokinase has been proposed to be a key component of a complex mechanism that promotes skin invasion by Streptococcus pyogenes . This study was designed to compare ska gene message and protein levels in wild-type M1 serotype isolate 1881 and a more invasive variant recovered from the spleen of a lethally infected mouse . M1 isolates selected for invasiveness demonstrated enhanced levels of active plasminogen activator activity in culture . This effect was due to a combination of increased expression of the ska gene and decreased expression of the speB gene . The speB gene product, SpeB, was found to efficiently degrade streptokinase in vitro.

J Clin Microbiol, 2004 Feb, 42(2), 919 - 22
Enterococcus cecorum empyema thoracis successfully treated with cefotaxime; Woo PC et al.; We report the first case of Enterococcus cecorum empyema thoracis and spontaneous bacterial peritonitis in a 44-year-old man with underlying cirrhosis . The patient responded to cefotaxime (MIC, 0.25 microg/ml) treatment and drainage of the empyema . Susceptibility of E . cecorum to expanded-spectrum cephalosporins could be due to its production of types of penicillin-binding proteins similar to those produced by Streptococcus species rather than to those produced by Enterococcus species (as predicted by phylogenetic analysis of the 16S rRNA gene sequences).

J Clin Microbiol, 2004 Feb, 42(2), 855 - 7
Prevalence of vancomycin-resistant enterococcus in prenatal screening cultures; Miller MB et al.; Recommendations for the perinatal treatment of women colonized with Streptococcus agalactiae include vancomycin prophylaxis for those with severe penicillin allergies and antibiotic-resistant organisms . Because of potential postpartum infections due to vancomycin-resistant enterococci (VRE) and the possible spread of vancomycin resistance, the prevalence of VRE in prenatal screening cultures was determined.

J Clin Microbiol, 2004 Feb, 42(2), 764 - 8
Molecular epidemiology of multiresistant Streptococcus pneumoniae with both erm(B)- and mef(A)-mediated macrolide resistance; Farrell DJ et al.; Of a total of 1043 macrolide-resistant Streptococcus pneumoniae isolates collected from 24 countries as part of PROTEKT 1999-2000, 71 isolates tested positive for both the mef(A) and erm(B) genes . Of 69 isolates subjected to further molecular investigations, all were resistant to tetracycline, 63 (91.3%) were resistant to penicillin, and 57 (82.6%) were resistant to trimethoprim-sulfamethoxazole . One isolate was also fluoroquinolone resistant, and another was resistant to quinupristin-dalfopristin . The ketolide telithromycin retained activity against all of the isolates . Of the 69 of these 71 isolates viable for further testing, 46 were from South Korea, 13 were from the United States, 8 came from Japan, and 1 each came from Mexico and Hungary . One major clonal complex (59 {85.5%} of 69 isolates) was identified by serotyping (with 85.5% of the isolates being 19A or 19F), pulsed-field gel electrophoresis, and multilocus sequence typing . The remaining isolates were less clonal in nature . Representative isolates were shown to carry the mobile genetic elements Tn1545 and mega, were negative for Tn1207.1, had tetracycline resistance mediated by tet(M), and contained the mef(E) variant of mef(A) . All isolates were positive for mel, a homologue of the msr(A) efflux gene . These clones are obviously very efficient at global dissemination, and hence it will be very important to monitor their progress through continued surveillance . Telithromycin demonstrated high levels of activity (MIC for 90% of the strains tested, 0.5 micro g/ml; MIC range, 0.06 to 1 micro g/ml) against all isolates.

Appl Environ Microbiol, 2004 Feb, 70(2), 929 - 36
Shifts in the membrane fatty acid profile of Streptococcus mutans enhance survival in acidic environments; Fozo EM et al.; Acid adaptation of Streptococcus mutans UA159 involves several different mechanisms, including the ability to alter its proportion of long-chain, monounsaturated membrane fatty acids (R . G . Quivey, Jr., R . Faustoferri, K . Monahan, and R . Marquis, FEMS Microbiol . Lett . 189:89-92, 2000) . In the present study, we examined the mechanism and timing of changes in fatty acid ratios and the potential benefit that an increased proportion of long-chained fatty acids has for the organism during growth at low pH . Cells taken from steady-state cultures at intermediate pH values of 6.5, 6, and 5.5 showed incremental changes from the short-chained, saturated membrane fatty acid profile normally seen in pH 7 cultures to the long-chained, monounsaturated fatty acids more typically observed in acidic cultures (pH 5) . Our observations showed that the bacterium was capable of effecting the majority of changes in approximately 20 min, far less than one generation time . However, reversion to the distribution of fatty acids seen in cells growing at a pH of 7 required a minimum of 10 generations . Fatty acid composition analysis of cells taken from cultures treated with chloramphenicol suggested that the changes in fatty acid distribution did not require de novo protein synthesis . Cells treated with the fatty acid biosynthesis inhibitor cerulenin were unable to alter their membrane fatty acid profiles and were unable to survive severe acidification . Results presented here indicate that membrane fatty acid redistribution is important for low pH survival and, as such, is a component of the S . mutans acid-adaptation arsenal.

Front Biosci, 2004 Jan 01, 9, 891 - 914
Extracellular virulence factors of Streptococcus pneumoniae; Jedrzejas MJ; Streptococcus pneumoniae is one of the major human bacterial pathogens . Current prophylactic agents against this pathogen are limited in their protective abilities and the role of therapeutics has been inadequate as resistant strains emerge . The development of new and improved therapies to combat the pneumococcal disease is necessary . In order to accomplish this, an understanding of the interactions between this bacterium and the host tissues is essential . Such interactions largely involve extracellular virulence factors that are expressed by the pathogen to interact with the host . These virulence factors include those based on sugars (glycome-based) as their building blocks, and proteins that are built from amino acids (proteome-based) . The first group includes primarily the capsule, teichoic and lipoteichoic acids . The second group is diverse and includes numerous surface proteins that are attached to the cell wall of pneumococci utilizing a variety of methods . For the purpose of this review these surface proteins were divided into three categories, proteins bound to peptidoglycan, those bound to choline residues present on the surface of penumococci, and those bound to the lipids of the cytoplasmic membrane . Both the glycome-based and protein-based virulence factors are described, analyzed, and represented graphically . Whenever possible, structural properties of these molecules were introduced.

J Heart Valve Dis, 2004 Jan, 13(1), 73 - 7
The Ross procedure as the surgical treatment of active aortic valve endocarditis; Birk E et al.; BACKGROUND AND AIM OF THE STUDY: The authors' experience is reported of aortic valve replacement (AVR) using the pulmonary autograft in patients with active aortic valve endocarditis, including an urgent Ross procedure in infants with the acute condition . METHODS: Nine patients aged between 8 months and 38 years, with a diagnosis of aortic valve endocarditis, have undergone AVR using the Ross procedure at the authors' institution since October 1997 . The diagnosis was established by clinical and echocardiographic findings . Indications for surgery were severe aortic insufficiency and congestive heart failure in all patients, with the addition of thromboembolic events (n = 3), persistent hyperpyrexia (n = 3) and vegetations (n = 5) . Four infants with no history of congenital cardiac malformation underwent urgent surgery because of acute bacterial endocarditis and rapid hemodynamic deterioration . Blood cultures were positive for Streptococcus pneumoniae in three patients, and Kingella kingi and Staphylococcus aureus in one patient each . Four patients were culture-negative . All patients were treated with intravenous antibiotics for four to six weeks postoperatively . RESULTS: There were no perioperative or late deaths, and no recurrent endocarditis at the implanted valves . Echocardiographic evaluation at discharge showed trivial to mild aortic insufficiency, with no stenosis at the left ventricular outflow tract . Similar findings were found across the right ventricular outflow tract . At follow up (range: 4 months to 5.5 years), none of the patients showed progression of aortic valve insufficiency or developed stenosis; three had mild and moderate homograft stenosis (Doppler gradient 20-40 mmHg), and all children had moderate homograft insufficiency . CONCLUSION: The Ross procedure is an excellent therapeutic option for active aortic valve endocarditis in young patients, and demonstrates low morbidity and mortality . Early surgery may be indicated in patients with acute aortic valve endocarditis because of the rapidly progressive nature of this disease.

J Dairy Sci, 2004 Jan, 87(1), 225 - 31
Effects of an automatic postmilking teat dipping system on new intramammary infections and iodine in milk; Galton DM; A technology of automatically applying a postmilking teat dip via the milking machine prior to machine detachment was compared to manual postmilking teat dipping with a teat dip cup for effects on new IMI and iodine content in milk . One hundred twenty Holstein cows were experimentally challenged in a 22-wk trial with Streptococcus agalactiae and Staphylococcus aureus and 148 Holstein cows were experimentally challenged with Streptococcus uberis in another 22-wk trial . The bacterial suspensions were applied to teats of all of the cows after premilking udder preparation and immediately prior to milking machine attachment . In both trials, cows were divided among four treatments: no postmilking teat dipping; manual postmilking teat dipping with a proven efficacious iodophor teat dip; manual postmilking teat dipping with an iodophor teat dip formulated for an automatic postmilking teat dipping system; and automatically postmilking teat dipping via milking machines with an iodophor teat dip formulated for the automatic postmilking teat dipping system . The postmilking teat dipping treatments reduced new Staph . aureus IMI by 64.5, 76.5, and 88.2%; new Strep . agalactiae IMI by 61.5, 77.8, and 94.4%; and new Strep . uberis IMI by 63.5, 82.5, and 93.8%, respectively, against the treatment of no postmilking teat dipping . The treatment applying the postmilking teat dip automatically via milking machines had the lowest number of new IMI caused by the three pathogens . Teat end and teat skin condition were characterized as normal at the end of the study with no differences between treatments . There were no differences with regard to iodine content in milk between treatments.

Clin Infect Dis, 2004 Feb 15, 38(4), 508 - 14 Epub 2004 Jan 28.
High-level penicillin-nonsusceptible Streptococcus pneumoniae bacteremia: identification of a low-risk subgroup; Ruhe JJ et al.; High-level penicillin resistance has been associated with treatment failure in patients with Streptococcus pneumoniae infections . To identify a subgroup of patients at low risk for high-level penicillin-nonsusceptible S . pneumoniae bacteremia, a cross-sectional study of 303 patients was performed . For the total study population, penicillin resistance was observed in 98 (32%) of 303 patients; high-level resistance was seen in 33 (11%) . A predictive model was created by using 3 baseline variables that were independently associated with high-level penicillin resistance: previous beta -lactam antibiotic use, previous stay in a risk area (defined as stay in day care facilities, prisons, homeless shelters, nursing homes, or other long-term care facilities), and previous respiratory tract infection . The model was used to identify patients at low and high risk for high-level penicillin-resistant pneumococcal bacteremia . None of the isolates of patients in the low-risk subgroup had ceftriaxone resistance . Patients in the low-risk subgroup could be empirically treated with fluoroquinolone-sparing regimens.

Mol Microbiol, 2004 Feb, 51(4), 1071 - 86
Interconnection of competence, stress and CiaR regulons in Streptococcus pneumoniae: competence triggers stationary phase autolysis of ciaR mutant cells; Dagkessamanskaia A et al.; Of the 13 two-component signal transduction systems (TCS) identified in Streptococcus pneumoniae, two, ComDE and CiaRH, are known to affect competence for natural genetic transformation . ComD and ComE act together with the comC-encoded competence-stimulating peptide (CSP) and with ComAB, the CSP-dedicated exporter, to co-ordinate activation of genes required for differentiation to competence . Several lines of evidence suggest that the CiaRH TCS and competence regulation are interconnected, including the observation that inactivation of the CiaR response regulator derepresses competence . However, the nature of the interconnection remains poorly understood . Interpretation of previous transcriptome analyses of ciaR mutants was complicated by competence derepression in the mutants . To circumvent this problem, we have used microarray analysis to investigate the transition from non-competence to competence in a comC-null wild-type strain and its ciaR derivative after the addition of CSP . This study increased the number of known CSP-induced genes from approximately 47 to 105 and revealed approximately 42 genes with reduced expression in competent cells . Induction of the CiaR regulon, as well as the entire HrcA and part of the CtsR stress response regulons, was observed in wild-type competent cells . Enhanced induction of stress response genes was detected in ciaR competent cells . In line with these observations, CSP was demonstrated to trigger growth arrest and stationary phase autolysis in ciaR cells . Taken together, these data strongly suggest that differentiation to competence imposes a temporary stress on cells, and that the CiaRH TCS is required for the cells to exit normally from the competent state.

Mol Microbiol, 2004 Feb, 51(4), 1051 - 70
Identification of competence pheromone responsive genes in Streptococcus pneumoniae by use of DNA microarrays; Peterson SN et al.; Natural genetic transformation in Streptococcus pneumoniae is controlled in part by a quorum-sensing system mediated by a peptide pheromone called competence-stimulating peptide (CSP), which acts to coordinate transient activation of genes required for competence . To characterize the transcriptional response and regulatory events occurring when cells are exposed to competence pheromone, we constructed DNA microarrays and analysed the temporal expression profiles of 1817 among the 2129 unique predicted open reading frames present in the S . pneumoniae TIGR4 genome (84%) . After CSP stimulation, responsive genes exhibited four temporally distinct expression profiles: early, late and delayed gene induction, and gene repression . At least eight early genes participate in competence regulation including comX, which encodes an alternative sigma factor . Late genes were dependent on ComX for CSP-induced expression, many playing important roles in transformation . Genes in the delayed class (third temporal wave) appear to be stress related . Genes repressed during the CSP response include ribosomal protein loci and other genes involved in protein synthesis . This study increased the number of identified CSP-responsive genes from approximately 40 to 188 . Given the relatively large number of induced genes (6% of the genome), it was of interest to determine which genes provide functions essential to transformation . Many of the induced loci were subjected to gene disruption mutagenesis, allowing us to establish that among 124 CSP-inducible genes, 67 were individually dispensable for transformation, whereas 23 were required for transformation.

Mol Microbiol, 2004 Feb, 51(4), 963 - 71
Structural studies of Streptococcus pneumoniae EPSP synthase in unliganded state, tetrahedral intermediate-bound state and S3P-GLP-bound state; Park H et al.; The shikimate pathway synthesizes aromatic amino acids and other essential metabolites that are necessary for bacteria, plants and fungi to survive . This pathway is not present in vertebrates and therefore represents an attractive target for antibacterial agents . We have successfully crystallized and solved the structure of unliganded, inhibitor-liganded and tetrahedral intermediate (TI)-liganded forms of Streptococcus pneumoniae EPSP synthase . The overall topology of the S . pneumoniae EPSP synthase is similar to that of the Escherichia coli EPSP synthase . In addition, the majority of residues responsible for ligand binding were conserved between the two proteins . TI-liganded structure provides absolute configuration of the C-2 atom from the F-PEP moiety of the enzyme-bound intermediate and also defines key residues responsible for the enzyme reaction . Comparison of the unliganded state and substrate-bound state of the enzyme provides insights into the structural mechanisms involved in dynamic events of ligand binding, domain movement and closure . This structural study of the pathogenic bacteria S . pneumoniae EPSP synthase with inhibitor and TI will provide invaluable information for the design of new-generation antibiotics.

Biotechnol Prog, 2004 Jan-Feb, 20(1), 289 - 98
Distribution of cells between solid/liquid and liquid/liquid interfaces; Bermudez O et al.; The use of aqueous two-phase systems (ATPSs) and each system's individual phase-forming species to prevent Streptococcus sanguis attachment onto hydroxyapatite discs was explored . The strategy that we followed was to attach the cells to a solid surface in the presence of an additional interface . Conditions under which, simultaneously, the phase-forming species form two phases and the cells proliferate were identified . Growth curves were constructed in the presence of various polymers and salts commonly used to prepare ATPSs . Several aqueous two-phase systems were selected such that bacterial growth was comparable to that observed in pure medium . Cells were allowed to attach to hydroxyapatite discs for 7 days in the presence of varying concentrations of media, media with polymer, media with salt, and media with ATPS . Streptococcus sanguis attachment to the disks was evaluated by scanning electron microscopy . The addition of a PEG/Na(2)SO(4) ATPS to high concentrations of yeast-tryptone (YT) media (>65%) and of a PEG/MgSO(4) ATPS to nutrient-limited media reduces surface coverage of S . sanguis to less than 10% . Comparison of the attachment levels for the systems containing PEG/Na(2)SO(4) to media containing the individual phase-forming species and to the YT reference systems indicated that nutrient availability did not affect attachment.

J Oral Sci, 2003 Dec, 45(4), 181 - 4
Detection of Streptococcus anginosus and 8-hydroxydeoxyguanosine in saliva; Sugano N et al.; Several studies have demonstrated a close association between Streptococcus (S.) anginosus infection and head and neck cancer . Accumulation of 8-hydroxy-deoxyguanosine (8-OHdG), which may result from the continuous generation of reactive oxygen species associated with chronic inflammation, has been reported in human preneoplastic lesions and in cancerous tissues . The purpose of the present investigation was to assess the salivary levels of S . anginosus and 8-OHdG in patients with periodontitis . Salivary levels of S . anginosus were measured by real-time PCR . S . anginosus was detected in 28 out of 38 (73.7%) of subjects . The 8-OHdG level was significantly higher in patients positive for S . anginosus than in patients negative for the bacterium . A significant decrease in S . anginosus and 8-OHdG levels was observed after initial periodontal treatment . Our findings indicate that, although the levels of S . anginosus are relatively low, there is a correlation between the salivary level of S . anginosus and 8-OHdG, and that periodontal treatment can decrease the levels of these hazard factors.

J Bacteriol, 2004 Feb, 186(4), 1106 - 9
ISSa4-based differentiation of Streptococcus agalactiae strains and identification of multiple target sites for ISSa4 insertions; Dmitriev A et al.; A collection of 113 epidemiologically unrelated Streptococcus agalactiae strains were studied (group B streptococcus; GBS): they belonged to different serotypes and were isolated from pregnant women in China and Russia . The insertion sequence ISSa4 was found in 21 of 113 strains (18,6%) . All of the strains with ISSa4 belonged to serotypes II and II/c and were characterized by the presence of IS1381 and IS861 as well as the absence of IS1548 and GBSi1 . All of the strains with ISSa4 possessed both bca and bac virulence genes coding for alpha and beta antigens, respectively . Among 21 ISSa4-positive strains, 13 different HindIII patterns (D1 to D13) hybridizing with an ISSa4 probe were found . One of them (D13) contained a single HindIII hybridization fragment 6.5 kb in size that was found to be specific for all ISSa4-positive GBS strains . Multiple target sites for insertions of ISSa4 were identified and included a putative pathogenicity island, "housekeeping" genes, and intergenic regions, as well as the genes for hypothetical proteins . No significant similarity was observed in the sequences of the target genes for ISSa4 insertions, in the relative location of the target genes on the chromosome, or the biological functions of the encoded proteins . The possible significance of ISSa4-based differentiation of the strains and the presence of possible "hot spots" for insertions of ISSa4 in GBS genome are discussed.

Expert Rev Vaccines, 2004 Feb, 3(1), 43 - 58
Lipid core peptide technology and group A streptococcal vaccine delivery; Olive C et al.; The antiphagocytic surface M protein of group A streptococcus has been widely studied as the major candidate antigen for a vaccine to prevent group A streptococcus infection . Approaches that have proven to be effective in animal models include the use of multi-epitope vaccines incorporating highly variable amino terminal serotypic determinants, those based on the carboxy terminal conserved region and combination vaccines incorporating both serotypic and conserved region determinants of the M protein . The use of lipid core peptide technology is at the forefront of this research in the quest to develop a broad-strain protective vaccine that can be delivered via the mucosal route, stimulating mucosal and systemic immunity . This review aims to cover the various strategies and technologies that have been investigated with regard to group A streptococcus vaccine design and development.

Curr Opin Pediatr, 2004 Feb, 16(1), 107 - 12
Current challenges in lower respiratory infections in children; Klig JE; PURPOSE OF REVIEW: Lower respiratory infections threaten the health of children worldwide . Streptococcus pneumoniae remains the most common bacterial cause of lower respiratory infection in children, whereas viral pathogens dominate as a more common cause of lower respiratory infection illness in infants and children overall . The diagnosis and clinical management of lower respiratory infections pose challenges to pediatric health providers as new technology is developed and new pathogens emerge in the spectrum of clinical disease . RECENT FINDINGS: Human metapneumovirus is now recognized as a cause of lower respiratory infection disease in children, and coronavirus has been linked to epidemics of severe acute respiratory syndrome . Respiratory syncytial virus continues to be a major source of viral lower respiratory infection illness in children and can lead to childhood asthma . Treatment for respiratory syncytial virus bronchiolitis depends largely on the severity of disease and the course of clinical symptoms . The diagnosis of bacterial lower respiratory infection disease remains a clinical challenge, but new methods to detect S . pneumoniae, or Chlamydia pneumoniae and Mycoplasma pneumoniae may facilitate the clinical management of these illnesses . As immunization against S . pneumoniae becomes more widely used, the complications of bacterial lower respiratory infections will diminish markedly . SUMMARY: Future progress in the clinical management of lower respiratory infection diseases will entail improved methods of early diagnosis, broader options for treatment, and better defined clinical parameters for triage and follow-up of children with lower respiratory infections.

Enferm Infecc Microbiol Clin, 2004 Jan, 22(1), 13 - 7
{Microbiological, clinical and epidemiological aspects of Streptococcus pneumoniae isolates recovered over two years}; Viciana MI et al.; INTRODUCTION: Streptococcus pneumoniae is the most frequent cause of non-hospital acquired pneumonia and meningitis in adults, and bacterial otitis media in children . Moreover, it causes a third of all acute sinusitis cases . Penicillin has been the treatment of choice for almost 50 years . Gradually, penicillin-resistant pneumococci have appeared throughout the world . Our aim was to investigate the epidemiology, pattern of resistance and serotypes of Streptococcus pneumoniae infection in our area . METHODS: Over a period of two years (May 1997-May 1999), Streptococcus pneumoniae strains were isolated in the Clinical Microbiology Unit of the University Hospital Virgen de la Victoria in Malaga, Spain . This is a 750-bed hospital covering a population of 407,480 inhabitants, and admitting 21,500 hospitalized patients per year . Streptococcus was identified by standard procedures: serotyping was done with the Quellung test and antibiotic susceptibility study by the disk diffusion method and E-test . RESULTS: Streptococcus pneumoniae infection was diagnosed in 170 patients during the years studied . The microorganism was isolated from samples of sputum (82), blood (43), aspirated bronchial fluid, cerebrospinal fluid (6), and exudates (7) . Non-hospital origin was identified in 88% of cases . The mean hospital stay was 12 days and mortality was 12.4% . Some 45.9% of the isolated strains were resistant to penicillin and 20% to cefotaxime . We found 31 different serotypes, with 77% of the isolated strains belonging to 12 serotypes . Serotypes 19, 3 and 6B were the most frequent in non-hospital infection, whereas 9V and 23F were related with nosocomial infection . Penicillin-resistant strains of Streptococcus pneumoniae belonged to 19 different serotypes; 6B, 9V, 14, 19 and 23F were the most important . CONCLUSIONS: As was expected, Streptococcus pneumoniae infections of mainly non-hospital origin in our area were characterized by elevated mortality and high-level resistance to penicillin . Immunosuppression was a predisposing factor.

Graefes Arch Clin Exp Ophthalmol, 2004 Mar, 242(3), 204 - 9 Epub 2004 Feb 04.
Epidemiological characteristics of microbiological results on patients with infectious corneal ulcers: a 13-year survey in Paraguay; Laspina F et al.; BACKGROUND: This is a retrospective, chart-reviewed study of patients diagnosed with infectious corneal ulcers at the Ophthalmology Department of the National University of Asuncion in Paraguay . The microbiological culture results are described, as well risk factors for the development of fungal keratitis . METHODS: After obtaining approval from the Institutional Review Board, an analysis of medical charts from 1988 to 2001 was conducted and 660 patients were identified to have been diagnosed with infectious corneal ulcers due to bacteria or fungi . Demographic data were recorded, including age, gender, occupation and geographic location of their home and work (city or rural) . Other information collected included the history of the presenting illness, past and current use of ocular medications and whether or not they had a history of trauma or contact lens use . Each patient had an eye examination performed by an ophthalmologist and corneal scrapings were obtained for cultures in all cases . Microbiologic culture results were analyzed . RESULTS: Twenty-one percent (136/660) of the specimens collected from the patients' conjunctiva and cornea were sterile in all culture media . Of the 524 (79%) positive cultures, 267 were due to bacteria (51%), 136 to fungi (26%), and 121 (23%) cultures yielded both fungi and bacteria . Of the 430 isolated bacteria approximately 25% (103) were coagulase negative Staphylococcus, followed by 23% (94) Staphylococcus aureus, 14% (60) Pseudomonas aeruginosa and 13% (56) Streptococcus pneumoniae . Acremonium species accounted for 40% (79) of all fungi identified, followed by Fusarium species (15%) (41) . Approximately two-thirds of the patients were male (n = 435) . For those patients for whom a history was available, approximately half had a history of trauma . Of these, half of these again involved foreign bodies . Over-the-counter medications were used commonly, and most of those patients had a delay in diagnosis of over 1 week . Risk factors for fungal keratitis as opposed to bacterial keratitis were male gender, agricultural occupation, age between 30 and 59, history of trauma and self-medication . CONCLUSIONS: The results of this study provide demographic data on patients with infectious corneal ulcers in Paraguay . Common causes of such ulcers are both bacteria and fungi . Most patients had self-medicated, and most had delayed seeking professional medical care.

Clin Microbiol Infect, 2004 Feb, 10(2), 177 - 81
Characterisation of invasive pneumococcal isolates in Catalan children up to 5 years of age, 1989-2000; Latorre C et al.; Ninety-six Streptococcus pneumoniae strains isolated between January 1989 and December 2000 from usually sterile sites of children aged < 5 years of age were included in the study . Resistance to penicillin (38.6% intermediate, 10.4% high-level), cefotaxime (20.8%), tetracycline (41.7%), chloramphenicol (33.3%) and erythromycin (27.1%), as well as serogroup/type, were related to age and pathology . Strains from children aged < 2 years showed the highest penicillin resistance rate . Resistance to penicillin, tetracycline, chloramphenicol and erythromycin was the most common pattern (18.8% of strains) . Most isolates (80.2%) belonged to serogroups/types included in the heptavalent conjugate vaccine.

Clin Microbiol Infect, 2004 Feb, 10(2), 174 - 7
Similar inflammatory response in human whole blood to live Streptococcus pneumoniae of different serotypes; Kragsbjerg P et al.; Differences in inflammatory responses in human adult whole blood to live pneumococcal serotypes 3, 7F, 9V and 23F were investigated . Using flow cytometry and ELISA, oxidative burst, expression of activation markers CD11b/CD18, and in-vitro production of tumour necrosis factor-alpha, interleukin-6 (IL-6) and interleukin-8 were measured . There was no significant difference between the serotypes regarding any of the variables investigated, although there was a trend towards higher concentrations of IL-6 induced by serotypes 9V and 23F . In the present experimental model, the serotypes of Streptococcus pneumoniae shown previously to cause different degrees of inflammation were found to cause a similar inflammatory response in human whole blood.

Clin Microbiol Infect, 2004 Apr, 10 Suppl 2, 28 - 35
Performance in practice: bacteriological efficacy in patients with drug-resistant S . pneumoniae; Garau J; Using pharmacokinetic/pharmacodynamic principles, pharmacokinetically enhanced amoxicillin/clavulanate 2000/125 mg twice daily was designed to provide adequate levels of amoxicillin over the 12-h dosing interval to eradicate penicillin-resistant Streptococcus pneumoniae (PRSP, penicillin MICs > or = 2 mg/L) with amoxicillin MICs of at least 4 mg/L . The clinical efficacy of amoxicillin/clavulanate 2000/125 mg was evaluated in patients with respiratory tract infections caused by S . pneumoniae, including isolates with elevated penicillin (2-8 mg/L) MICs . Data from 10 clinical studies were combined: seven randomised (1:1), double-blind, controlled trials (efficacy intent-to-treat {ITT}N = 3376): amoxicillin/clavulanate 2000/125 mg twice daily vs . levofloxacin 500 mg once daily in acute bacterial sinusitis (ABS); levofloxacin 500 mg once daily in acute exacerbations of chronic bronchitis (AECB); clarithromycin 500 mg twice daily in AECB; amoxicillin/clavulanate 875/125 mg twice daily/three times daily and 1000/125 mg three times daily in community-acquired pneumonia (CAP) and three noncomparative studies (efficacy ITT N = 3024): two in ABS, one in CAP . The bacteriological per-protocol (PP) population at follow up (days 14-39) comprised 1295 patients for amoxicillin/clavulanate 2000/125 mg and 241 for comparators . With amoxicillin/clavulanate 2000/125 mg at follow-up, outcome was successful (clinical success and eradication/presumed eradication) in 85/90 (94.4%) patients with S . pneumoniae in comparative studies and 421/445 (94.6%) in noncomparative studies, and with comparators 58/70 (82.9%) were successes . In the amoxicillin/clavulanate 2000/125 mg group at follow up, 52/552 S . pneumoniae isolates were resistant to penicillin . At follow up, 50/52 (96.2%) patients with PRSP were successes, including 6/7 with amoxicillin MICs of 4 mg/L and 7/8 with amoxicillin MICs of 8 mg/L . Success rates for amoxicillin/clavulanate 2000/125 mg against PRSP were similar for CAP (96.0%{24/25}), AECB (100%{3/3}) and ABS (95.8%{23/24}) . There were six PRSP isolates in the comparator group (two isolates were from one patient), and three of five patients in this group were successes . In conclusion, amoxicillin/clavulanate 2000/125 mg demonstrated combined clinical/bacteriological success against 50/52 patients with PRSP, including 13/15 strains with amoxicillin MICs of 4-8 mg/L . These results for the pharmacokinetic-enhanced formulation of amoxicillin/clavulanate 2000/125 mg are in line with the high efficacy against PRSP predicted using pharmacokinetic/pharmacodynamic parameters.

Clin Microbiol Infect, 2004 Apr, 10 Suppl 2, 12 - 7
Proof of concept: performance testing in models; Craig WA; Pharmacokinetic (PK) and pharmacodynamic (PD) principles that predict antimicrobial efficacy can be used to set targets for antimicrobial design and optimisation . Although current formulations of amoxicillin and amoxicillin/clavulanate have retained their efficacy against many, but not all, penicillin-nonsusceptible Streptococcus pneumoniae, additional coverage is required to address the growing problem of drug-resistant strains . Accordingly, two new oral formulations of amoxicillin/clavulanate, a paediatric formulation at 90/6.4 mg/kg/day and a pharmacokinetically enhanced formulation at 2000/125 mg twice daily for adults, were designed using PK/PD principles . These principles indicate that for amoxicillin and amoxicillin/clavulanate, a time above MIC of 35-40% of the dosing interval is predictive of high bacterial efficacy . In line with PK/PD predictions, simulation of human pharmacokinetics in in-vitro kinetic models and in a rat model of pneumonia, amoxicillin/clavulanate 2000/125 mg twice daily was highly effective against S . pneumoniae strains with amoxicillin MICs of 4 or 8 mg/L . Against strains with amoxicillin MICs of 4 mg/L, amoxicillin/clavulanate 2000/125 mg twice daily was significantly more effective than the conventional 875/125 mg twice daily formulation, azithromycin and levofloxacin, even though all levofloxacin MICs were < or = 1 mg/L . Following infection with S . pneumoniae strains with amoxicillin MICs of 8 mg/L, the amoxicillin/clavulanate 2000/125 mg twice daily formulation was more effective than the conventional amoxicillin/clavulanate formulations of 875/125 mg twice daily and three times daily and 1000/125 mg three times daily, and had similar or better efficacy than azithromycin and levofloxacin, depending on the strain . These data indicate the potential benefit of therapy with amoxicillin/clavulanate 2000/125 mg twice daily compared with conventional formulations and other marketed antimicrobials in the treatment of respiratory tract infection.

Laryngoscope, 2004 Feb, 114(2), 273 - 8
Experimental sinusitis in a rhinogenic model; Kara CO et al.; OBJECTIVES/HYPOTHESIS: The objectives were to determine the optimal sinusitis induction period and to examine microbiological and histopathological changes of sinusitis recovery stage in a rhinogenic sinusitis model . METHODS: A synthetic sponge was inserted into the right-side nasal cavities of rabbits . The sponge was impregnated with a Streptococcus pneumoniae strain in group 1 and with sterile saline solution in groups 2 and 3 . After the fourth day of sponge insertion, sinuses were examined by coronal computed tomography scans at two-day intervals until any radiological evidence of sinusitis was observed . When sinusitis was detected radiologically, five rabbits each from groups 1 and 2 were killed for histological examination . To determine the recovery period of sinusitis, sponges were removed from the rest of the rabbits in groups 1 and 2 . Rabbits were selected randomly and killed on the 15th and the 30th days of the recovery period, immediately after radiological examinations . Group 3 was considered a sham group . RESULTS: Sinusitis induction was performed in all rabbits in groups 1 and 2 until the 8th day . After the sponges were removed, inflammation persisted until the 30th day of the study . CONCLUSION: In a rhinogenic sinusitis model, although histological features of sinusitis were demonstrated, further studies are required to standardize this model and to examine whether or not the studied bacterial strain spreads from nasal cavity into sinus.

Bioinformatics, 2004 Mar 22, 20(5), 790 - 7 Epub 2004 Jan 29.
CAAT-Box, Contigs-Assembly and Annotation Tool-Box for genome sequencing projects; Frangeul L et al.; MOTIVATION: Contigs-Assembly and Annotation Tool-Box (CAAT-Box) is a software package developed for the computational part of a genome project where the sequence is obtained by a shotgun strategy . CAAT-Box contains new tools to predict links between contigs by using similarity searches with other whole genome sequences . Most importantly, it allows annotation of a genome to commence during the finishing phase using a gene-oriented strategy . For this purpose, CAAT-Box creates an Individual Protein file (IPF) for each ORF of an assembly . The nucleotide sequence reported in an IPF corresponds to the sequence of the ORF with 500 additional bases before the ORF and 200 bases after . For annotation, additional information like Blast results can be added or linked to the IPFs as well as automatic and/or manual annotations . When a new assembly is performed, CAAT-Box creates new IPFs according to the old IPF panel . CAAT-Box recognizes the modified IPFs which are the only ones used for a new automatic analysis after each assembly . Using this strategy, the user works with a group of IPFs independently of the closure phase progression . The IPFs are accessible by a web server and can therefore be modified and commented by different groups . RESULT: CAAT-Box was used to obtain and to annotate several complete genomes like Listeria monocytogenes or Streptococcus agalactiae . AVAILABILITY: The program may be obtained from the authors and is freely available to non-profit organisations.

Surg Neurol, 2004 Feb, 61(2), 142 - 3; discussion 143-4
Atypical presentation of spontaneous discitis: case report; Han L et al.; BACKGROUND: Spontaneous discitis is primarily a pediatric illness . Adult patients usually present at an average age of 69 years with a history of diabetes or with a systemic infection . The lumbar spine is the most frequent site of infection (54%), and the cervical is the least at 10% . The causative organisms are most commonly Staphylococcus aureus and beta-hemolytic streptococcus species . Intravenous antibiotics are the mainstays of treatment, and surgical intervention is usually not required . CASE PRESENTATION: A single case observation with an unusual presentation from the statistically typical criterion of discitis is described . CONCLUSIONS: Atypical discitis needs to be considered in the differential diagnoses in the middle-aged and healthy population.

J La State Med Soc, 2003 Nov-Dec, 155(6), 325 - 31
Education does pay off: pneumococcal vaccine screening and administration in hospitalized adult patients with pneumonia; Kruspe R et al.; Streptococcus pneumoniae-associated infections are an important cause of hospitalization and mortality in high-risk and elderly patients . Even in the setting of appropriate therapy, the case fatality rate of invasive pneumococcal disease in the elderly may approach 40% . Since approximately 40,000 people die annually from pneumococcal-associated disease, it represents a substantial target for vaccine-preventable, bacterial fatalities . The 23-valent pneumococcal polysaccharide vaccine has proven consistently effective in preventing invasive pneumococcal disease . Despite its endorsement by numerous specialty societies, the pneumococcal vaccine is underutilized in the inpatient setting . In a recent report of quality indicators for Medicare beneficiaries, the percentage of Medicare beneficiaries in Louisiana admitted with pneumonia who were screened or received the pneumococcal vaccination prior to discharge was only 4%, the lowest percentage in the United States . The Louisiana State University-New Orleans Internal Medicine Department and its house staff embarked upon a retrospective study to determine its baseline pneumococcal vaccination or screening rates for all patients with pneumonia on its inpatient services at the The Medical Center of Louisiana in New Orleans from July 2000 through June 2001 . From July 2001 through June 2002 an intensive educational intervention concentrating on the indications and benefits of pneumococcal vaccination was directed toward the Louisiana State University Internal Medicine house staff assigned to the inpatient service . Retrospective analysis for pneumococcal vaccine screening and administration of charts of all patients with pneumonia on the LSU Medicine service from July 2001 through June 2002 was performed in order to determine the effects of the intervention . Data from the pre-educational intervention period revealed a baseline pneumococcal vaccine screening or administration rate of 11% for all patients with pneumonia on the LSU Internal Medicine inpatient service . During the one-year intervention period, the pneumococcal vaccine screening or administration rate increased to 71%, a clinically and statistically significant increase (p-value < 0.0001) . Data targeting patients 65 years of age and older revealed a baseline pneumococcal vaccine screening or administration rate of 10% for patients with pneumonia on the LSU Internal Medicine inpatient service which increased to 82% during the one year educational intervention (p-value < 0.0001) . House officer scores (possible range 0-100) on a questionnaire assessing their understanding of the indications and benefits of pneumococcal vaccination were significantly higher after the educational intervention compared to before the intervention (means +/- standard deviations, 68 +/- 9 vs . 59 +/- 10, p < 0.0001) . The findings from this study highlight the importance of education in increasing compliance with widely-accepted practice guidelines such as pneumococcal vaccine screening or administration in patients hospitalized with pneumonia.

Indian J Med Res, 2003 Jun, 117, 247 - 52
Rapid diagnosis of vaginal carriage of group B beta haemolytic streptococcus by an enrichment cum antigen detection test; Das A et al.; BACKGROUND & OBJECTIVES: Group B beta haemolytic streptococcus (GBS) is a frequent colonizer of the maternal genital tract causing peripartum fever, puerperal sepsis, neonatal sepsis and neonatal meningitis . The conventional methods for detection of maternal colonization take 24-48 h . We made an attempt to standardize a rapid enrichment cum antigen detection test to screen pregnant women for GBS colonization in less than 8 h, so as to enable early institution of measures to prevent neonatal sepsis . METHODS: Vaginal swabs of 100 women >36 wk of gestation were inoculated onto enrichment broth (Todd Hewitt broth with lysed horse blood and antibiotics) . After incubation for 1,2,4,6, and 18 h, the broth was cultured on sheep blood agar . In culture positive cases, the enrichment broth was subjected to antigen detection by latex agglutination test (LAT) . For further evaluation of the rapid test, another group of 100 pregnant women were screened for GBS carriage by 6 h enrichment broth culture followed by antigen detection test . RESULTS: Five of the first group yielded GBS on culture and all were positive for GBS antigen after 6 h enrichment . Thirteen of the second group were positive for the antigen, but GBS could be isolated in ten only . This enrichment cum antigen detection test showed sensitivity, specificity, and positive and negative predictive values of 100, 98.4, 83.3 and 100 per cent respectively and could detect as few as 10(3) cfu/ml organisms . Maternal vaginal carriage of GBS was 7.5 per cent (15/200) . INTERPRETATION & CONCLUSION: Six hours of enrichment followed by antigen detection proved to be a rapid and reliable method for detection of GBS colonization . This test is easy to perform making it an ideal test for screening GBS vaginal colonization at labour and starting chemoprophylaxis, where indicated on the same day, before the woman is discharged.

Rev Med Liege, 2003 Nov, 58(11), 675 - 80
{Problematic resistance of pneumococcal antibiotic resistance}; Marchal V et al.; Streptococcus pneumoniae is the most common cause of community-acquired pneumonia and it is also a common cause of sinusitis, otitis media, bacteremia and meningitis . The increasing resistance to antimicrobial agents, now endemic in many countries, reflects an uncontrolled use of antibiotics . A good antibiotics policy and vaccination are at the moment the only way to control efficaciously the increasing antibiotic resistance of Streptococcus pneumoniae.

Acta Crystallogr D Biol Crystallogr, 2004 Feb, 60(Pt 2), 359 - 61 Epub 2004 Jan 23.
Expression, purification, crystallization and preliminary characterization of uridine 5'-diphospho-N-acetylmuramoyl L-alanyl-D-glutamate:lysine ligase (MurE) from Streptococcus pneumoniae 110K/70; Blewett AM et al.; An ORF designated sp1530 (murE) in the Streptococcus pneumoniae TIGR4 genome sequence, identified as uridine 5'-diphospho-N-acetylmuramoyl-L-alanyl-D-glutamate:L-lysine ligase (MurE; EC 6.3.2.7), was cloned into the high-expression plasmid pET21b and overexpressed in Escherichia coli BL21 (DE3) Star . The enzyme was purified in three steps to 99% purity . Crystals were obtained by the hanging-drop vapour-diffusion method at 291 K from solutions containing 25%(w/v) polyethylene glycol 2000 monomethylether, 0.2 M potassium thiocyanate, 0.1 M MES pH 6.5 in the presence of uridine 5'-diphospho-N-acetylmuramoyl alanyl glutamate (UDP-MurNAc-L-Ala-D-Glu) with and without 5'-adenylyl imidophosphate (AMP-PNP), a non-hydrolysable analogue of ATP . Diffraction data to 1.5 and 2.7 A, respectively, were collected at the European Synchrotron Radiation Facility (ESRF) . Crystals grown in the presence of two ligands belong to space group P1, with unit-cell parameters a = 68.4, b = 71.4, c = 74.8 A, alpha = 73.4, beta = 80.5, gamma = 72.3 degrees . Crystals grown in the presence of UDP-MurNAc-L-Ala-D-Glu alone belong to space group P2(1), with unit-cell parameters a = 71.1, b = 129.4, c = 74.6 A, beta = 106.3 degrees.

J Infect Dis, 2004 Feb 1, 189(3), 385 - 92 Epub 2004 Jan 16.
Four antibiotic-resistant Streptococcus pneumoniae clones unrelated to the pneumococcal conjugate vaccine serotypes, including 2 new serotypes, causing acute otitis media in southern Israel; Porat N et al.; This study examined the prevalence of antibiotic-resistant clones that belong to serotypes not included in the pneumococcal conjugate vaccines and that cause a significant percentage of acute otitis media (AOM) in children in southern Israel . During 1998-2001, 2467 pneumococcal isolates, obtained from middle-ear fluid of children <3 years old with AOM, were characterized by antimicrobial susceptibility testing, serotype testing, and pulsed-field gel electrophoresis . Non-vaccine type (NVT) strains constituted 477 (19%) of the 2467 isolates, of which 173 (36%) belonged to only 4 serotypes: 35B, 33F, 21, and 15B/C . For serotype 35B, 47 (96%) of 49 strains were penicillin nonsusceptible, and 93% constituted a single clone; for serotype 33F, 31 (82%) of 38 strains were penicillin nonsusceptible, and 95% constituted a single clone; for serotype 21, 38 (93%) of 41 strains were penicillin nonsusceptible, and 93% constituted a single clone; for serotype 15B/C, 22 (49%) of 45 strains were penicillin nonsusceptible, and 42% constituted a single clone . Two of these clones have not been described elsewhere . The high prevalence of NVT clones should increase the awareness of the potential for replacement of the vaccine strains with these NVT antibiotic-resistant strains.

Biochemistry, 2004 Feb 3, 43(4), 1065 - 74
Rates of elementary catalytic steps for different metal forms of the family II pyrophosphatase from Streptococcus gordonii; Zyryanov AB et al.; Soluble inorganic pyrophosphatases (PPases) form two nonhomologous families, denoted I and II, that have similar active-site structures but different catalytic activities and metal cofactor specificities . Family II PPases, which are often found in pathogenic bacteria, are more active than family I PPases, and their best cofactor is Mn(2+) rather than Mg(2+), the preferred cofactor of family I PPases . Here, we present results of a detailed kinetic analysis of a family II PPase from Streptococcus gordonii (sgPPase), which was undertaken to elucidate the factors underlying the different properties of family I and II PPases . We measured rates of PP(i) hydrolysis, PP(i) synthesis, and P(i)/water oxygen exchange catalyzed by sgPPase with Mn(2+), Mg(2+), or Co(2+) in the high-affinity metal-binding site and Mg(2+) in the other sites, as well as the binding affinities for several active-site ligands (metal cofactors, fluoride, and P(i)) . On the basis of these data, we deduced a minimal four-step kinetic scheme and evaluated microscopic rate constants for all eight relevant reaction steps . Comparison of these results with those obtained previously for the well-known family I PPase from Saccharomyces cerevisiae (Y-PPase) led to the following conclusions: (a) catalysis by sgPPase does not involve the enzyme-PP(i) complex isomerization known to occur in family I PPases; (b) the values of k(cat) for the magnesium forms of sgPPase and Y-PPase are similar because of similar rates of bound PP(i) hydrolysis and product release; (c) the marked acceleration of sgPPase catalysis in the presence of Mn(2+) and Co(2+) results from a combined effect of these ions on bound PP(i) hydrolysis and P(i) release; (d) sgPPase exhibits lower affinity for both PP(i) and P(i); and (e) sgPPase and Y-PPase exhibit similar values of k(cat)/K(m), which characterizes the PPase efficiency in vivo (i.e., at nonsaturating PP(i) concentrations).

Pediatr Infect Dis J, 2004 Jan, 23(1), 56 - 61
Molecular characterization of Streptococcus pyogenes isolates collected during periods of increased acute rheumatic fever activity in Utah; Miner LJ et al.; BACKGROUND: Salt Lake City, Utah has seen a continuing resurgence of rheumatic fever (RF) since 1985 . METHODS: emm genotyping and multilocus sequence typing of streptococcal isolates from periods of increased RF activity were performed . RESULTS: Multiple genotypes were present during 1985 and 1998, two peak years of RF activity, and in 1992, a year with reduced RF activity . emm3 and emm18.1 were present in 1985 and 1998, but not in 1992 . Two other emm types, 12 and L28, were significantly elevated in 1998 (a peak RF year) over 1992 (a non-peak RF year) . Allelic profiles for the emm3 and emm18.1 isolates exhibited clonality . CONCLUSIONS: During years of increased RF activity multiple emm types, including emm18.1 and emm3, were circulating in the community . During a year of decreased RF activity, emm3 and emm18.1 genotypes were absent . The clonality of the emm3 and emm18.1 types suggests that specific clones of both types are important in the resurgence of RF during these peak years . Two other genotypes, emm12 and emmL28, may also be associated with the persistence of RF in the Salt Lake City, UT area.

Pediatr Infect Dis J, 2004 Jan, 23(1), 47 - 52
Dexamethasone effects on group B streptococcal infection in newborn rats; Tran TV et al.; BACKGROUND: We previously published that human neutrophil-mediated bacterial killing of group B Streptococcus (GBS) in vitro was dependent on the timing and concentration of dexamethasone exposure . HYPOTHESIS: Dexamethasone treatment would affect neutrophil mediated killing of GBS in an animal model . METHODS: Wistar rat pups were randomly allocated to receive placebo or dexamethasone before, early or late after GBS infection . Suckling rats were infected with 104 or 105 colony-forming units of GBS or nothing . Pups were followed for survival, quantitative bacteremia, growth and neutrophil-mediated bacterial killing . Neutrophils for bacterial killing were obtained via cardiac puncture before infection . Statistics included chi square for survival, Mann-Whitney U test for bacteremia, analysis of variance for growth and paired Student's t test for bacterial killing analyses . RESULTS: Dexamethasone treatment before invasive GBS infection decreases quantitative bacteremia, improves survival and improves neonatal neutrophil-mediated bacterial killing in suckling rats, whereas dexamethasone treatment after infection increases bacteremia and decreases survival . Regardless of timing of dexamethasone treatment, before or after invasive GBS infection, growth was significantly impaired in all suckling rats receiving dexamethasone compared with controls . CONCLUSION: Treatment with dexamethasone before invasive GBS infection improves survival and decreases bacteremia in suckling rats; this appears in part to be mediated by improved neonatal neutrophil-mediated bacterial killing . We speculate that this improvement in outcome may be a result of improved number or function of neutrophil cell surface receptors.

Infect Immun, 2004 Feb, 72(2), 1192 - 4
Hydrogen peroxide-mediated killing of Caenorhabditis elegans: a common feature of different streptococcal species; Bolm M et al.; Recently, we reported that Streptococcus pyogenes kills Caenorhabditis elegans by the use of hydrogen peroxide (H2O2) . Here we show that diverse streptococcal species cause death of C . elegans larvae in proportion to the level of H2O2 produced . H2O2 may mask the effects of other pathogenicity factors of catalase-negative bacteria in the C . elegans infection model.

Infect Immun, 2004 Feb, 72(2), 1184 - 7
Intranasal immunization of mice with group B streptococcal protein rib and cholera toxin B subunit confers protection against lethal infection; Larsson C et al.; Intranasal immunization of mice with Rib, a cell surface protein of group B streptococcus (GBS), conjugated to or simply coadministered with the recombinant cholera toxin B subunit, induces systemic immunoglobulin G (IgG) and local IgA antibody responses and confers protection against lethal GBS infection . These findings have implications for the development of a human GBS vaccine.

Infect Immun, 2004 Feb, 72(2), 818 - 23
Host-pathogen interaction during pneumococcal infection in patients with chronic obstructive pulmonary disease; Bogaert D et al.; Acute exacerbation is a frequent complication of chronic obstructive pulmonary disease (COPD) . Recent studies suggested a role for bacteria such as Streptococcus pneumoniae in the development of acute exacerbation . For this study, we investigated the following in COPD patients: (i) the epidemiology of pneumococcal colonization and infection, (ii) the effect of pneumococcal colonization on the development of exacerbation, and (iii) the immunological response against S . pneumoniae . We cultured sputa of 269 COPD patients during a stable state and during exacerbation of COPD and characterized 115 pneumococcal isolates by use of serotyping . Moreover, we studied serum immunoglobulin G (IgG) antibody titers, antibody avidities, and functional antibody titers against the seven conjugate vaccine serotypes in these patients . Colonization with only pneumococci (monocultures) increased the risk of exacerbation, with a hazard ratio of 2.93 (95% confidence interval, 1.41 to 6.07) . The most prevalent pneumococcal serotypes found were serotypes 19F, 3, 14, 9L/N/V, 23A/B, and 11 . We calculated the theoretical coverage for the 7- and 11-valent pneumococcal vaccines to be 60 and 73%, respectively . All patients had detectable IgG levels against the seven conjugate vaccine serotypes . These antibody titers were significantly lower than those in vaccinated healthy adults . Finally, on average, a 2.5-fold rise in serotype-specific and functional antibodies in S . pneumoniae-positive sputum cultures was observed during exacerbation . Our data indicate that pneumococcal colonization in COPD patients is frequently caused by vaccine serotype strains . Moreover, pneumococcal colonization is a risk factor for exacerbation of COPD . Finally, our findings demonstrate that COPD patients are able to mount a significant immune response to pneumococcal infection . COPD patients may therefore benefit from pneumococcal vaccination.

Infect Immun, 2004 Feb, 72(2), 788 - 94
Role of Toll-like receptor 4 in gram-positive and gram-negative pneumonia in mice; Branger J et al.; To determine the role of Toll-like receptor 4 (TLR4) in the immune response to pneumonia, C3H/HeJ mice (which display a mutant nonfunctional TLR4) and C3H/HeN wild-type mice were intranasally infected with either Streptococcus pneumoniae (a common gram-positive respiratory pathogen) or Klebsiella pneumoniae (a common gram-negative respiratory pathogen) . In cases of pneumococcal pneumonia, TLR4 mutant mice showed a reduced survival only after infection with low-level bacterial doses, which was associated with a higher bacterial burden in their lungs 48 h postinfection . In Klebsiella pneumonia, TLR4 mutant mice demonstrated a shortened survival after infection with either a low- or a high-level bacterial dose together with an enhanced bacterial outgrowth in their lungs . These data suggest that TLR4 contributes to a protective immune response in both pneumococcal and Klebsiella pneumonia and that its role is more important in respiratory tract infection caused by the latter (gram-negative) pathogen.

Infect Immun, 2004 Feb, 72(2), 774 - 81
Investigation of a novel DNase of Streptococcus suis serotype 2; Fontaine MC et al.; A secreted nuclease, SsnA, was identified in the virulent Streptococcus suis isolate SX332 and subsequently in each of the type strains of capsular serotypes 1 through 9 . Screening of 258 porcine clinical isolates from surface (nasal mucosa or palatine tonsil) or internal (joint, brain or other internal organ) locations revealed a significant relationship (P < 0.001) between expression of nuclease and isolation from an internal site . A 3,126-bp gene, ssnA, was identified from a phenotypically nuclease-negative pGh9:ISS1 insertion mutant, and analysis of the predicted SsnA sequence revealed a 35-amino-acid (aa) secretion signal sequence, a 22-aa DNA-binding domain, and a typical gram-positive cell wall sorting motif . A requirement of Ca2+ and Mg2+ for SsnA activity was determined, and the substrate specificity was found to be for single- and double-stranded linear DNA . Reverse transcription-PCR experiments revealed that ssnA is expressed throughout all stages of S . suis growth, and Western blots with porcine anti-S . suis immune sera against a recombinant, truncated SsnA derivative (rSsnADelta) confirmed that SsnA is expressed in vivo . Furthermore, anti-rSsnADelta antibodies were sufficient to neutralize SsnA activity . Analyses of subcellular fractions of SX332 and derived mutants, on DNA-containing polyacrylamide gels and by Western blotting, suggest that SsnA is cell wall located.

Infect Immun, 2004 Feb, 72(2), 623 - 8
Contribution of CsrR-regulated virulence factors to the progress and outcome of murine skin infections by Streptococcus pyogenes; Engleberg NC et al.; Streptococcus pyogenes with null mutations in the csrRS regulatory locus are highly virulent in mice due to derepression of hyaluronic acid capsule synthesis and exotoxins, e.g., streptolysin S (SLS) and pyrogenic exotoxin B (SpeB) . We generated derivatives of a DeltacsrRS strain that also carry deletions in hasAB (leading to an acapsular phenotype) or in sagA (phenotypically SLS-) or an interruption of speB (SpeB-) to test the relative contributions of these factors to the development of necrotic skin lesions . Inoculation of 2 x 10(6) to 4 x 10(6) CFU of either acapsular or SLS- strains into hairless mice resulted in lesions approximately 70% smaller than those of the DeltacsrRS parent strain . Elimination of SLS also reduced lethality from 100% to 0% at this inoculum (P < 10(-7); Fisher exact test) . In contrast, SLS+ SpeB- mutants yielded lesions that were only 41% smaller than the parent strain (t = 2.2; P = 0.04), but only 3 the 17 lesions had dermal sloughing (P = 10(-5)) . The nonulcerative lesions associated with SpeB- strains appeared pale with surrounding erythema . We conclude that capsule and SLS contribute to the subcutaneous spread of S . pyogenes and to a fatal outcome of infection . SpeB facilitates early dermal ulceration but has minor influence on lesion size and mortality . Large ulcerative lesions are observed only when both toxins are present.

Br J Anaesth, 2004 Mar, 92(3), 427 - 9 Epub 2004 Jan 22.
Fatal streptococcal necrotizing fasciitis as a complication of axillary brachial plexus block; Nseir S et al.; A 74-yr-old diabetic woman developed necrotizing fasciitis of the right upper limb after axillary brachial plexus block for carpal tunnel decompression . Clinical signs included oedema, diffuse swelling and bullae; rapidly followed by toxic shock syndrome and multiorgan failure . The patient died 48 h after hospital admission, despite broad-spectrum antibiotics, surgical treatment and supportive measures for the management of shock and multiorgan failure . Cultures yielded group A Streptococcus . Delay in antibiotic and surgical treatment probably affected the outcome . Early diagnosis and treatment are essential to improve the outcome of streptococcal necrotizing fasciitis.

Carcinogenesis, 2004 Aug, 25(8), 1477 - 84 Epub 2004 Jan 23.
Carcinogenic properties of proteins with pro-inflammatory activity from Streptococcus infantarius (formerly S.bovis); Biarc J et al.; Several studies reported linkage between bacterial infections and carcinogenesis . Streptococcus bovis was traditionally considered as a lower grade pathogen frequently involved in bacteremia and endocarditis . This bacterium became important in human health as it was shown that 25-80% of patients who presented a S.bovis bacteremia had also a colorectal tumor . Moreover, in previous experiments, we demonstrated that S.bovis or S.bovis wall extracted antigens (WEA) were able to promote carcinogenesis in rats . The aim of the present study was: (i) to identify the S.bovis proteins responsible for in vitro pro-inflammatory properties; (ii) to purify them; (iii) to examine their ability to stimulate in vitro IL-8 and COX-2 expression by human colon cancer cells; and (iv) to assess in vivo their pro-carcinogenic potential in a rat model of colon carcinogenesis . The purified S300 fraction, as determined by proteomic analysis, contained 72 protein spots in two-dimensional gel electrophoresis representing 12 different proteins able to trigger human epithelial colonic Caco-2 cells and rat colonic mucosa to release CXC chemokines (human IL-8 or rat CINC/GRO) and prostaglandins E2, correlated with an in vitro over-expression of COX-2 . Moreover, these proteins were highly effective in the promotion of pre-neoplastic lesions in azoxymethane-treated rats . In the presence of these proteins, Caco-2 cells exhibited enhanced phosphorylation of the three classes of MAP kinases . Our results show a relationship between the pro-inflammatory potential of S.bovis proteins and their pro-carcinogenic properties, confirming the linkage between inflammation and colon carcinogenesis . These data support the hypothesis that colonic bacteria can contribute to cancer development particularly in chronic infection/inflammation diseases where bacterial components may interfere with cell function.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 605 - 7
Activities of cethromycin and telithromycin against recent North American isolates of Streptococcus pneumoniae; Jorgensen JH et al.; The in vitro activities of two investigational ketolides, cethromycin (formerly ABT-773) and telithromycin, were determined for a selected group of 312 Streptococcus pneumoniae isolates from a national surveillance program . The MIC of cethromycin at which 50% of the isolates were inhibited was 0.008 micro g/ml, and the MIC at which 90% of the isolates were inhibited was 0.06 micro g/ml; the corresponding values for telithromycin were </=0.015 and 0.25 micro g/ml, respectively . For six quinupristin-dalfopristin-resistant strains, the cethromycin MICs were 0.25 to 16 micro g/ml and the telithromycin MICs were 1 to 4 micro g/ml . However, there was only 0.3% resistance to telithromycin.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 378 - 83
Pharmacodynamics of S-3578, a novel cephem, in murine lung and systemic infection models; Miyazaki S et al.; S-3578 is a novel beta-lactam with enhanced activity against drug-resistant gram-positive cocci such as methicillin-resistant Staphylococcus aureus (MRSA) . We used murine penicillin-resistant Streptococcus pneumoniae lung infection and neutropenic murine systemic MRSA infection models to determine the pharmacokinetic (PK)-pharmacodynamic (PD) parameter that best correlated with efficacy . Pharmacokinetic studies revealed that the maximum concentration in serum/dose values for S-3578 and cefepime in plasma in the lung infection model were 1.21 to 1.54 and 0.97 to 1.29, respectively; those for S-3578 in plasma in the systemic infection model were 0.78 to 1.02 . The area under the concentration-time curve (AUC)/dose values for S-3578 and cefepime in plasma in the lung infection model were 0.98 to 1.13 and 0.77 to 1.04, respectively, and those for S-3578 in plasma in the systemic infection model were 1.03 to 1.11 . The half-lives of S-3578 and cefepime in plasma in the lung infection model were 0.29 to 0.38 and 0.29 to 0.34, respectively, and those of S-3578 in plasma in the systemic infection model were 0.40 to 0.61 . The time above the MIC was the PK-PD parameter that best correlated with efficacy in the murine lung infection model (R(2) = 84 and 92% for S-3578 and cefepime in plasma, respectively) . There was a twofold increase in the dose of S-3578 in the systemic infection model compared to that in the pneumonia model, yet the AUCs were the same . This may be due to the different MICs for the two pathogens.

J Neuroimmunol, 2004 Feb, 147(1-2), 33 - 4
Pneumococcus virulence factor sialidase: a new direction in neuro-AIDS research?
Rosenberg A.
The purpose of this presentation is to invite consideration by the research community of the hypothesis that sialidase, a virulence factor of Streptococcus pneumoniae (pneumococcus) and most other opportunistic co-infectious agents associated with HIV infection, advances progression of HIV infection to neuro-AIDS.

J Immune Based Ther Vaccines . 2004 Jan 23;2(1):2.
Development of a model of focal pneumococcal pneumonia in young rats; Malley R et al.; BACKGROUND: A recently licensed pneumococcal conjugate vaccine has been shown to be highly effective in the prevention of bacteremia in immunized children but the degree of protection against pneumonia has been difficult to determine . METHODS: We sought to develop a model of Streptococcus pneumoniae pneumonia in Sprague-Dawley rats . We challenged three-week old Sprague-Dawley pups via intrapulmonary injection of S . pneumoniae serotypes 3 and 6B . Outcomes included bacteremia, mortality as well histologic sections of the lungs . RESULTS: Pneumonia was reliably produced in animals receiving either 10 or 100 cfu of type 3 pneumococci, with 30% and 50% mortality respectively . Similarly, with type 6B, the likelihood of pneumonia increased with the inoculum, as did the mortality rate . Prophylactic administration of a preparation of high-titered anticapsular antibody prevented the development of type 3 pneumonia and death . CONCLUSION: We propose that this model may be useful for the evaluation of vaccines for the prevention of pneumococcal pneumonia.

Microbes Infect, 2004 Jan, 6(1), 86 - 92
Phytoestrogen genistein as an anti-staphylococcal agent; Verdrengh M et al.; The soybean-derived isoflavone genistein has been shown to exert beneficial effects on many disorders, including cancer and cardiovascular diseases . The effects of genistein on mammalian cells are mediated by its abilities to inhibit topoisomerase II and protein tyrosine kinase . In order to examine the potential antibacterial activities of genistein, we incubated the bacteria with various concentrations of this compound for different periods of time and assessed the viable counts . Exposure to genistein exhibited an inhibitory effect on all staphylococcal strains tested, including methicillin-resistant strains . Furthermore, the growth of Streptococcus pasteurianus, Bacillus cereus, and Helicobacter pylori was clearly inhibited by genistein, whereas Escherichia coli growth was not suppressed . Daidzein, which is structurally similar to genistein, but deficient in topoisomerase II inhibitory activity, also inhibited the growth of Staphylococcus aureus, albeit with lower potency than genistein . Our results indicate that genistein exerts potent antibacterial properties in vitro, which are possibly mediated by the stabilization of the covalent topoisomerase II-DNA cleavage complex.

Curr Drug Targets, 2004 Jan, 5(1), 57 - 69
Vaccine development for group A streptococcus infections and associated disease; Batzloff MR et al.; Group A streptococcus (GAS) is responsible for a number of diseases ranging from uncomplicated pharyngitis through to life-treating invasive and post-infectious diseases such as necrotizing fasciitis and rheumatic heart disease . GAS associated diseases occur globally and are serious problems in many developing nations and indigenous populations of many developed nations . This, and the resurgence in industrialized countries, and increased virulence of GAS in the 1980s highlight the need of cost-effective control strategies . Here we highlight the GAS diseases that are still a problem in many populations and discuss potentially useful strategies to combat GAS infections and disease.

Ann Vasc Surg, 2003 Sep, 17(5), 554 - 7
Multiple mycotic aortic aneurysms due to Streptococcus pneumoniae; Coutu M et al.; Primary pneumococcal aortic mycotic aneurysms are rare clinical entities . Only a few cases have been reported in the literature . Extremely rare presentation is the occurrence of three simultaneous aneurysms . Treatment usually necessitates intravenous antibiotherapy combined with staged surgical interventions . This report highlights the case of a 52-year-old man with multiple Streptococcus pneumoniae mycotic aneurysms that were simultaneously and successfully treated during a one-stage surgical procedure . The aorta may be prone to infection, especially when its intima is structurally altered by pathologic processes like atherosclerosis, inflammation or trauma . Mycotic aneurysm is a rare but serious vascular condition needing urgent medical and surgical attention because of potential lethal complications.

J Child Neurol, 2003 Dec, 18(12), 828 - 34
Regulation of brain-derived neurotrophic factor (BDNF) expression following antibiotic treatment of experimental bacterial meningitis; Li L et al.; Although more and more new potent antibiotics have been used, mortality and neurologic deficits still occur frequently following bacterial meningitis in children . In this article, the expression of brain-derived neurotrophic factor messenger ribonucleic acid (RNA) and its production in the brains of rats were investigated during the course of experimental bacterial meningitis and after treatment with an antibiotic plus dexamethasone . In the brains of Streptococcus pneumoniae-inoculated rats, brain-derived neurotrophic factor (BDNF) messenger RNA was obviously up-regulated after inoculation for 24 hours (P < .01) and then declined but was still greater than that in the brains of control rats after inoculation for 5 days (P < .05) . The expression of brain-derived neurotrophic factor in the brains of infected rats treated by antibiotic was dose dependent, down-regulated, and almost undetectable (P < .01) but up-regulated after treatment with an antibiotic plus dexamethasone (P < .01) . However, the expression of brain-derived neurotrophic factor messenger RNA did not change in control rats treated with an antibiotic . Brain-derived neurotrophic factor protein showed similar changes, except it declined to normal levels 5 days after inoculation . Brain-derived neurotrophic factor messenger RNA and its production were observed in some infiltrating inflammatory cells in the brain of infected rats . The results of our studies support the hypothesis that brain-derived neurotrophic factor might play a neuroprotective role in brain damage during bacterial meningitis, and the expression of brain-derived neurotrophic factor messenger RNA and its production might be inhibited after treatment with antibiotics . The findings suggest that both eradicating the bacterial pathogen with antibiotics and adjuvant administering of brain-derived neurotrophic factor might be more beneficial to prevent brain damage.

Infection, 2003 Dec, 31(6), 425 - 7
Meningitis due to multiple-resistant penicillin- and cefotaxime-intermediate Streptococcus pneumoniae in a German child after bone marrow transplantation; Buxmann H et al.; The incidence of infection with penicillin-non-susceptible Streptococcus pneumoniae is increasing rapidly worldwide . Spain and France are highly affected, whereas the level of penicillin resistance in Germany, Italy, The Netherlands and Scandinavia is low . We report a lethal episode of meningitis due to penicillin- and cefotaxime-intermediate S . pneumoniae in a 7-year-old, allogene bone marrow transplanted German boy, 5 weeks after a holiday in Spain . Three days prior to the infection the patient showed good performance status . He was in complete remission without signs of graft-versus-host disease (GVHD) . He died on day 341 post bone marrow transplant (BMT), 2 days after the onset of meningitis . Penicillin-non-susceptible S . pneumoniae should be regarded as a potential infectious agent even in countries with a low prevalence of resistance.

Mol Biotechnol, 2004 Jan, 26(1), 81 - 6
Method for introducing specific and unmarked mutations into the chromosome of Streptococcus pneumoniae; Iannelli F et al.; This work describes a procedure for the generation of site-specific mutations into the chromosome of Streptococcus pneumoniae that does not involve the use of an antibiotic resistance marker . A linear fragment of transforming deoxyribonucleic acid (DNA) is constructed by polymerase chain reaction (PCR) (gene splicing by overlap extension) and used to transform competent cells of S . pneumoniae . Selection of transformants is performed by PCR, and typically, 1% of the transformed cells show the expected mutation . By this protocol it is possible to change a single base pair into the pneumococcal genome, as well as obtaining in-frame deletions and insertions.

J Biol Chem, 2004 Apr 16, 279(16), 16463 - 70 Epub 2004 Jan 20.
A PBP2x from a clinical isolate of Streptococcus pneumoniae exhibits an alternative mechanism for reduction of susceptibility to beta-lactam antibiotics; Pernot L et al.; The human pathogen Streptococcus pneumoniae is one of the main causative agents of respiratory tract infections . At present, clinical isolates of S . pneumoniae often exhibit decreased susceptibility toward beta-lactams, a phenomenon linked to multiple mutations within the penicillin-binding proteins (PBPs) . PBP2x, one of the six PBPs of S . pneumoniae, is the first target to be modified under antibiotic pressure . By comparing 89 S . pneumoniae PBP2x sequences from clinical and public data bases, we have identified one major group of sequences from drug-sensitive strains as well as two distinct groups from drug-resistant strains . The first group includes proteins that display high similarity to PBP2x from the well characterized resistant strain Sp328 . The second group includes sequences in which a signature mutation, Q552E, is found adjacent to the third catalytic motif . In this work, a PBP2x from a representative strain from the latter group (S . pneumoniae 5259) was biochemically and structurally characterized . Phenotypical analyses of transformed pneumococci show that the Q552E substitution is responsible for most of the reduction of strain susceptibility toward beta-lactams . The crystal structure of 5259-PBP2x reveals a change in polarity and charge distribution around the active site cavity, as well as rearrangement of strand beta3, emulating structural changes observed for other PBPs that confer drug resistance to Gram-positive pathogens . Interestingly, the active site of 5259-PBP2x is in closed conformation, whereas that of Sp328-PBP2x is open . Consequently, S . pneumoniae has evolved to employ the same protein in two distinct mechanisms of antibiotic resistance.

Curr Infect Dis Rep, 2004 Feb, 6(1), 7 - 12
What Is the Clinical Impact of Macrolide Resistance?
Lonks JR.
Respiratory tract infections are treated empirically . Treatment is based on the likely pathogens and their antibiotic susceptibility . The most common respiratory tract pathogen is Streptococcus pneumoniae . In the United States, approximately 25% to 30% of S . pneumoniae are resistant to erythromycin and other macrolides . There are two mechanisms of resistance: ribosomal methylation that causes high-level resistance, and an efflux pump that causes low-level resistance . Macrolides are ineffective in animal models that use pneumococcal isolates with the methylase- or efflux-mediated resistance mechanisms . There are many case reports that describe clinical failure and isolation of a macrolide-resistant pneumococcus while a patient receives macrolide treatment . Two recent studies that included macrolide-susceptible and macrolide-resistant pneumococci showed that breakthrough bacteremia in patients receiving macrolide treatment occurred only with macrolide-resistant isolates . Study of bacteremic disease ensures the pathogenic role of the pneumococcus; however, it underestimates the true clinical impact of macrolide resistance.

Zhonghua Er Ke Za Zhi, 2003 Sep, 41(9), 688 - 91
{Analysis of antimicrobial resistance of Streptococcus pneumoniae with restriction fragment length polymorphism of pbp2b gene and pulsed-field gel electrophoresis profiles among children}; Yu SJ et al.; Streptococcus pneumoniae is a common cause of potentially life-threatening infections such as meningitis, bacteraemia, pneumonia worldwide, for which children of preschool age are at particularly high risk . Since the late 1970s and 1980s, antibiotic resistance among pneumococci has become an emerging problem . Several multidrug-resistant clones have rapidly spread throughout the world . OBJECTIVE: (1) To investigate the prevalence of penicillin and other antibiotics nonsusceptibility among pneumococci . (2) To analyze the correlation of pbp2b amplicon profiles with penicillin resistance . (3) To serotype 31 isolates of penicillin-resistant pneumococci by latex agglutination . (4) To analyze the chromosomal relatedness of serotype 23F and 6 isolates of penicillin-resistant pneumococci by using pulsed-field gel electrophoresis (PFGE) and characterize these isolates in molecular epidemiology . METHODS: (1) Susceptibility was determined by using broth microdilution, E-test, and K-B disk . (2) The correlation of pbp2b amplicon profiles with penicillin resistance was assessed by restriction fragment length polymorphism (RFLP) . (3) Serotyping of penicillin-resistant pneumococcal isolates was performed by using latex agglutination . (4) The properties of serotype 23F and 6 isolates of penicillin-resistant pneumococci were assessed by PFGE . RESULTS: S . pneumoniae with increased nonsusceptibility (including intermediate strains and resistant strains) to penicillin G was 9.9% in 1997, 12.6% in 1998, 14.6% in 2000; to cefuroxime 4.2%, 1.5%, 8.2%; to cefotaxime 0.0%, 1.7%, 1.0% respectively . There were no statistically significant differences (P > 0.05) . While resistance to erythromycin, trimethoprim-sulfamethoxazole and chloramphenicol increased significantly from 76.8% in 1997 to 87.4% in 2000, from 74.7% to 88.3%, and from 22.6% to 40.8%, respectively (P < 0.05) . RFLP analysis of pneumococcal pbp2b-specific amplicons was effective for screening penicillin resistance . Of the 31 strains of penicillin-resistant pneumococci (MICs 0.12 - 2.0 micro g/ml) studied, 6 (19.4%) strains (MICs 0.12 - 0.19 micro g/ml) were serotype 23F and 3 (9.7%) strains (MICs 0.5 - 1.5 micro g/ml) were serotype 6 . There were nearly identical susceptibility to antibiotics and identical PFGE patterns in the former, and there were different susceptibility to antibiotics and different PFGE patterns in the latter . Three serotype 6 strains had different susceptibility to antibiotics and different PFGE patterns, which suggested that those strains may be scattered . CONCLUSION: Generally beta-lactams retained their activity against S . pneumoniae in Beijing . Resistance to erythromycin, trimethoprim-sulfamethoxazole, and chloramphenicol increased drastically . RFLP analysis of pneumococcal pbp2b-specific amplicons was effective for screening penicillin resistance . In 6 strains of serotype 23 F there were nearly identical susceptibility to antibiotics and identical PFGE patterns, which suggested the probability that there was a spread of serotype 23F isolates with low-level penicillin resistance in local area.

Int J Antimicrob Agents, 2004 Jan, 23(1), 32 - 8
Streptococcus pneumoniae in Saudi Arabia: antibiotic resistance and serotypes of recent clinical isolates; Memish ZA et al.; During 2000, 154 clinical Streptococcus pneumoniae isolates were collected from or through three major hospitals serving the Western, Central, and Eastern regions of the Kingdom of Saudi Arabia . Ninety-one (59.1%) of the 154 isolates were resistant to penicillin with 23 strains (14.9%) highly resistant . Resistance was more prevalent among isolates obtained from patients <10 years of age and from isolates cultured from blood and cerebral spinal fluid . Decreased sensitivity to ceftriaxone, erythromycin, trimethoprim-sulphamethoxazole, and levofloxacin was found in 14.9, 15.6, 9.7, and 1.3% of isolates, respectively . There were no significant differences in the resistance pattern between isolates obtained from patients in the three different geographical areas . Serotypes of 116 isolates revealed the most prevalent types to be (descending order) 4, 3, 19F, 9V, 6A, 19A, 14 and 23F, comprising 75% of all strains typed; 9.5% of isolates were nontypable . S . pneumoniae isolates from Saudi Arabia have become more resistant to penicillin and other antimicrobials over the past 20 years.

J AAPOS, 2003 Dec, 7(6), 413 - 7
Intracranial infection associated with preseptal and orbital cellulitis in the pediatric patient; Reynolds DJ et al.; PURPOSE: To identify risk factors in children admitted with preseptal or orbital cellulitis with associated intracranial infection . METHODS: A retrospective chart review identified 10 patients (< or = 18 years) with a diagnosis of preseptal or orbital cellulitis and a concurrent or subsequent diagnosis of intracranial infection . RESULTS: Diagnoses confirmed by imaging included sinusitis (n = 10), preseptal cellulitis (n = 4), orbital cellulitis (n = 6), orbital subperiosteal abscess (n = 5), Pott's puffy tumor (n = 4), epidural empyema (n = 2), epidural abscess (n = 6), and brain abscess (n = 2) . The timing of diagnosis of intracranial infection ranged from hospital day 1 to 21 . All but 1 patient had positive microbial cultures . Seven of 10 patients had positive microbial cultures from two or more sites, 70% of which were polymicrobial; Streptococcus species and Staphylococcus species were the most commonly isolated bacterial pathogens . All patients required both medical and surgical therapy; all 10 patients underwent sinus surgery; 8 patients required neurosurgical craniotomy; and 5 patients underwent orbital surgery . There were no deaths . CONCLUSION: Intracranial involvement should be suspected in any patient age > or = 7 years with preseptal or orbital cellulitis associated with orbital subperiosteal abscess, Pott's puffy tumor, concurrent sinusitis, complaints of headache, and continuing fever despite intravenous antibiotics . Given the high incidence of polymicrobial infection found on cultures in this series, broad-spectrum antibiotics are strongly indicated . When imaging the orbits and sinuses in such patients, we recommend including the brain to rule out intracranial involvement.

Pediatr Infect Dis J, 2004 Jan, 23(1 Suppl), S87 - 97
Respiratory viruses predisposing to bacterial infections: role of neuraminidase; Peltola VT et al.; BACKGROUND AND METHODS: Viral-bacterial coinfections in humans are well-documented . Viral infections often lead to bacterial superinfections . In vitro and animal models for influenza, as well as molecular microbiology study of viruses and bacteria, provide an understanding of the mechanisms that explain how respiratory viruses and bacteria combine to cause disease . This article focuses on viral and bacterial combinations, particularly synergism between influenza and Streptococcus pneumoniae . RESULTS: Potential mechanisms for synergism between viruses and bacteria include: virus destruction of respiratory epithelium may increase bacterial adhesion; virus-induced immunosuppression may cause bacterial superinfections; and inflammatory response to viral infection may up-regulate expression of molecules that bacteria utilize as receptors . Influenza and parainfluenza viruses possess neuraminidase (NA) activity, which appears to increase bacterial adherence after viral preincubation . Experimental studies demonstrate that viral NA exposes pneumococcal receptors on host cells by removing terminal sialic acids . Other studies show that inhibition of viral NA activity reduces adherence and invasion of S . pneumoniae, independently of effects on viral replication . Clinical studies reveal that influenza vaccination reduces the incidence of secondary bacterial respiratory tract infections . CONCLUSIONS: Detection of viral factors (e.g . high NA activity) that increase the likely potential of epidemic/pandemic influenza strains for causing morbidity and mortality from secondary bacterial infections provides new possibilities for intervention . Additional study is needed to identify the mechanisms for the development of bacterial complications after infections with respiratory syncytial virus and other important respiratory viruses that lack NA activity . Prevention of bacterial superinfection is likely to depend on effective antiviral measures.

Biosci Biotechnol Biochem, 2003 Dec, 67(12), 2567 - 73
Anti-cariogenic properties of a water-soluble extract from cacao; Ito K et al.; The addition of a water-soluble extract from cacao-extracted powder (CEPWS) to a cariogenic model food, a white chocolate-like diet that contains 35% sucrose, significantly reduced caries scores in SPF rats infected with Streptococcus sobrinus 6715, compared to control rats fed a white chocolate-like diet . CEPWS markedly inhibited water-insoluble glucan (WIG) synthesis through crude glucosyltransferases (GTFs) from Streptococcus sobrinus B13N in vitro . GTF-inhibitor(s) in CEPWS was prepared through three-step fractionation, and was termed CEPWS-BT, which is a high molecular weight (>10 kDa) heat-stable matrix of sugar, protein, and polyphenol . When the inhibitory effect of CEPWS-BT on glucan synthesis was examined using the purified GTF-I, GTF-T, and GTF-U enzymes from S . sobrinus B13N, significant reduction in GTF-I and GTF-T activity as a result of adding CEPWS-BT at low concentrations was observed . These results suggest that the addition of CEPWS to cariogenic food could be useful in controlling dental caries.

J Antimicrob Chemother, 2004 Feb, 53(2), 375 - 8 Epub 2004 Jan 16.
Mutations of the rpoB gene in rifampicin-resistant Streptococcus pneumoniae in Taiwan; Chen JY et al.; OBJECTIVE: To determine the mechanism of rifampicin resistance in Streptococcus pneumoniae in Taiwan . METHODS: Rifampicin resistance was investigated with respect to the rpoB gene in 23 invasive S . pneumoniae isolates collected from 1996 to 2001 . PCR and molecular typing were used for genetic and epidemiological analyses . Transformation was used to determine the functional gene for resistance . RESULTS: Twenty-two of 23 isolates carried at least one mutation at either cluster I or III of rpoB; the most frequent mutation found in 21 isolates (91%) was histidine (H499) to asparagine or tyrosine at position 499, followed by isoleucine to valine (I624V) at position 624 in 16 isolates (70%), tyrosine to phenylalanine (Y589F) at position 589 in 14 isolates (60.9%) and isoleucine to valine (I608V) at position 608 in 13 isolates (56.5%) . Less-frequent mutations were also identified: D489V, R597F, N623E, N623S, N669D, Q671K, Y674F and A683V . High-level rifampicin resistance was observed in isolates with a mutation at position 499 or 489 . Mutations other than at position 499 or 489 played little role in or had no relation to rifampicin resistance . No dominant epidemic strain was observed with ribotyping, multilocus sequence typing, or serotyping . CONCLUSIONS: Rifampicin resistance among multidrug-resistant S . pneumoniae in Taiwan was mostly caused by rpoB mutations.

J Bacteriol, 2004 Feb, 186(3), 706 - 12
The NeuC protein of Escherichia coli K1 is a UDP N-acetylglucosamine 2-epimerase; Vann WF et al.; The K1 capsule is an essential virulence determinant of Escherichia coli strains that cause meningitis in neonates . Biosynthesis and transport of the capsule, an alpha-2,8-linked polymer of sialic acid, are encoded by the 17-kb kps gene cluster . We deleted neuC, a K1 gene implicated in sialic acid synthesis, from the chromosome of EV36, a K-12-K1 hybrid, by allelic exchange . Exogenously added sialic acid restored capsule expression to the deletion strain (DeltaneuC), confirming that NeuC is necessary for sialic acid synthesis . The deduced amino acid sequence of NeuC showed similarities to those of UDP-N-acetylglucosamine (GlcNAc) 2-epimerases from both prokaryotes and eukaryotes . The NeuC homologue from serotype III Streptococcus agalactiae complements DeltaneuC . We cloned the neuC gene into an intein expression vector to facilitate purification . We demonstrated by paper chromatography that the purified neuC gene product catalyzed the formation of {2-(14)C}acetamidoglucal and {N-(14)C}acetylmannosamine (ManNAc) from UDP-{(14)C}GlcNAc . The formation of reaction intermediate 2-acetamidoglucal with the concomitant release of UDP was confirmed by proton and phosphorus nuclear magnetic resonance spectroscopy . NeuC could not use GlcNAc as a substrate . These data suggest that neuC encodes an epimerase that catalyzes the formation of ManNAc from UDP-GlcNAc via a 2-acetamidoglucal intermediate . The unexpected release of the glucal intermediate and the extremely low rate of ManNAc formation likely were a result of the in vitro assay conditions, in which a key regulatory molecule or protein was absent.

J Bacteriol, 2004 Feb, 186(3), 638 - 45
The Streptococcus gordonii platelet binding protein GspB undergoes glycosylation independently of export; Bensing BA et al.; The binding of bacteria and platelets may play a central role in the pathogenesis of infective endocarditis . Platelet binding by Streptococcus gordonii strain M99 is predominantly mediated by the 286-kDa cell wall-anchored protein GspB . This unusually large protein lacks a typical amino-terminal signal peptide and is translocated from the cytoplasm via a dedicated transport system . A 14-kb segment just downstream of gspB encodes SecA2 and SecY2, two components of the GspB-specific transport system . The downstream segment also encodes several putative glycosyl transferases that may be responsible for the posttranslational modification of GspB . In this study, we compared the abilities of M99 and two GspB(-) mutant strains to bind various lectins . GspB was found to have affinity for lectins that bind N-acetylglucosamine . We also examined variant forms of GspB that lack a carboxy-terminal cell wall-anchoring domain and thus are free of covalent linkage to cell wall peptidoglycan . Like native GspB, these truncated proteins appear to be heavily glycosylated, as evidenced by migration during sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an apparent molecular mass >100 kDa in excess of the predicted mass, negligible staining with conventional protein stains, and reactivity with hydrazide following periodate oxidation . Furthermore, analysis of the carbohydrate associated with the GspB variants by high-pH anion-exchange chromatography revealed the presence of approximately 70 to 100 monosaccharide residues per GspB polypeptide (primarily N-acetylglucosamine and glucose) . Analysis of GspB in protoplasts of secA2 or secY2 mutant strains, which do not export GspB, indicates that GspB is glycosylated in the cytoplasm of these strains . The combined data suggest that the native GspB is a glycoprotein and that it may be glycosylated prior to export.

AMIA Annu Symp Proc . 2003;:480-3.
Database-driven computerized antibiotic decision support: novel use of expert antibiotic susceptibility rules embedded in a pathogen-antibiotic logic matrix; Mullett CJ et al.; To better serve an antibiotic guidance program, we hypothesized that the relatively few antibiotic susceptibility measurements conducted in the microbiology laboratory could be extended to predict antibiotic susceptibilities for all antibiotics on the hospital formulary using expert infectious disease logic . With the assistance of infectious disease specialists, we developed these logic rules and then applied them to 26,196 unique patient culture specimens and the accompanying 334,131 antibiotic susceptibility measurements generating 804,809 additional predicted bug-drug susceptibility data points . From the resulting data set, the antibiotic susceptibility profile for one pathogen, Streptococcus pneumoniae, is highlighted herein . We then incorporated the extended susceptibility profiles into a computerized antibiotic guidance program that matches current patients of interest with the positive cultures from past similar patients and calculates predicted effective antibiotic therapy . We conclude that this method successfully derives antibiotic predictions and merits further testing to evaluate its potential use in the hospital environment.

Schweiz Arch Tierheilkd, 2003 Dec, 145(12), 571 - 5
{Current data on antibiotic resistance of the most important bovine mastitis pathogens in Switzerland}; Corti S et al.; 100 strains of Staphylococcus aureus (S . aureus), 100 strains of coagulase-negative staphylococci (CNS), 100 strains of Streptococcus spp . and 100 strains of Escherichia coli (E . coli), isolated from bovine mastitis milk samples between November 2002 and April 2003, were tested for their sensitivity to various antibiotics by means of the agar diffusion method . The antibiotics were chosen on the basis of their licenses for intramammary application in Switzerland . 91% of the S . aureus strains were sensitive to all the antibiotics tested . Only 9% of the strains were resistant to Penicillin G and 7% to Ampicillin . 53% of the CNS strains were sensitive to all the antibiotics tested . 31% exhibited resistance to Penicillin G, 26% to Ampicillin, 16% to Cloxacillin and 14% to Lincomycin . 30% of the Streptococcus spp . strains were sensitive to all the antibiotics tested . 4% were resistant to Penicillin G, 4% to Amoxicillin/clavulanic acid, 1% to Cefoperazone, 2% to Cefquinome, 35% to Neomycin, 22% to Gentamicin, 61% to Kanamy-cin and 11% to Lincomycin . 43% of the strains showed multiple resistance . 79% of the E . coli strains were sensitive to all the antibiotics tested . 20% exhibited resistance to Ampicillin, 9% to Neomycin and 10% to Kanamycin . A comparison of the own results with data of other authors in Switzerland shows no important changes in the resistance situation during the last 20 years . With the exception of two strains (Streptococcus spp.), all tested isolates were sensible against Cefquinome.

Otol Neurotol, 2004 Jan, 25(1), 14 - 8
Eustachian tube gland changes in acute otitis media; Caye-Thomasen P et al.; OBJECTIVES: To investigate the histomorphologic impact of acute otitis media on the subepithelial glands of the eustachian tube . Previous investigations have shown an increase of eustachian tube goblet cell density during and up to at least 6 months after acute otitis media . STUDY DESIGN: Longitudinal study in an experimental animal model of acute otitis media caused by Streptococcus pneumoniae . METHODS: Bacterial middle ear challenge, after which the animals were killed, in groups of five rats on Days 4, 8, 16, 90, and 180 . Dissection and preparation of the eustachian tube and adjacent structures, followed by serial transverse sectioning, periodic acid-Schiff/alcian blue staining, and qualitative/quantitative morphometric light-microscopic investigations of the histomorphology of the eustachian tube glands, in particular, the stainability, composition, volume, and number of gland acini . RESULTS: The volume of the eustachian tube glands progressed to peak 16 days after inoculation, followed by a gradual normalization . The volume was still increased 3 months after the acute infection but completely normalized after 6 months . The increase was primarily due to hypertrophy of the mucous gland components, as the serous gland component volume and number of gland acini was unchanged . The periodic acid-Schiff/alcian blue staining of the mucous gland acini changed temporarily during the acute infection, indicating changes in mucus glycoprotein composition, from neutral to acidic/sulfated . CONCLUSION: The volume of the eustachian tube glands increases during and up to at least 3 months after acute otitis media, primarily because of hypertrophy of the mucous gland components . This may compromise tubal ventilatory and drainage function.

Otol Neurotol, 2004 Jan, 25(1), 9 - 13
The efficacy of Burow's solution as an ear preparation for the treatment of chronic ear infections; Kashiwamura M et al.; OBJECTIVE: To determine the efficacy of Burow's solution as an otologic preparation for the treatment of chronic ear infection . STUDY DESIGN: Two studies were included: 1) a prospective clinical study and 2) a laboratory study on antibacterial and antifungal effects . SETTING: A private otology practice and a laboratory study . PATIENTS: Fifty-eight patients with refractory otorrhea . INTERVENTION: Diagnosis by otoscopy, audiometry, and bacteriology . METHODS: Burow's solution was mixed in solutions with four organisms: methicillin-resistant Staphylococcus aureus, penicillin-resistant Streptococcus pneumoniae, Candida albicans, and Aspergillus . Soon after Burow's solution was mixed in the solutions with organisms, and 5, 10, and 20 minutes thereafter, the mixtures were cultured on agars . The numbers of the bacterial or fungal colonies were counted to evaluate the effect of Burow's solution . MAIN OUTCOME MEASURES: Changes in the clinical findings of the ears, the symptom of otorrhea, and side effects were assessed . RESULTS: Thirty-five (70%) of the 50 ears assessed were "cured" and 10 (20%) ears assessed were "improved." No significant side effect was observed . Regarding the laboratory study, the four organisms disappeared within 20 minutes after Burow's solution was mixed . CONCLUSION: Burow's solution was considered to be an effective otologic preparation.

Scand J Infect Dis, 2003, 35(11-12), 878 - 81
Group B streptococcal necrotizing fasciitis and toxic shock-like syndrome: a case report and review of the literature; Crum NF et al.; We report the fourth case of group B streptococcal (GBS) necrotizing fasciitis and toxic shock-like syndrome . Since the mechanism of GBS toxic shock may be similar to Group A Streptococcus, intravenous immunoglobulin should be considered as an adjunct to clindamycin-based antibiotic therapy.

Scand J Infect Dis, 2003, 35(11-12), 876 - 8
Streptococcus pyogenes meningitis complicating varicella in a 3-month-old child; Brandt CM et al.; Varicella is a common, usually self-limited infectious disease, and complications are believed to be rare . Despite the dramatic increase in invasive Streptococcus pyogenes infections associated with varicella zoster virus infections in recent years, post-varicella S . pyogenes meningitis occurs very rarely . The third case in the literature is described here.

Scand J Infect Dis, 2003, 35(11-12), 843 - 50
Antibiotic consumption, bacterial resistance and their correlation in a Swiss university hospital and its adult intensive care units; Loeffler JM et al.; Ecological surveys of high-antibiotic use areas in the hospital should be used to evaluate patterns and trends in order to optimise antibiotic consumption and minimise resistance . We conducted a retrospective cohort study with the aim to examine trends in antimicrobial consumption and bacterial susceptibility at the Geneva University Hospital and its adult ICUs between 1996 and 2000 . The average annual consumption of antimicrobials was 400 d-defined doses (DDD)/1000 patient-d in the entire hospital, 462 in the surgical ICU and 683 in the medical ICU . In the medical ICU, we observed a steady decrease of intravenous antimicrobial use, whereas a 25% increase in the total antimicrobial consumption was noted in 1999 and 2000 for the entire hospital . The proportion of different bacterial species, resistance rates and antibiotic consumption rates differed between the entire hospital and the ICUs, with moderate variation between y . Possible relationships between antibiotic consumption and resistance rates, expressed as DDD and as number of resistant isolates per 1000 patient-d respectively, were calculated for of the most frequently isolated bacteria (total 71 correlations) . Statistically significant positive correlations between antibiotic consumption and resistance were found in Escherichia coli for piperacillin, in Pseudomonas aeruginosa for piperacillin, cephalosporins and aminoglycosides, in Klebsiella spp . for cephalosporin, in coagulase-negative staphylococci for gentamicin and in Streptococcus pneumoniae for penicillin.

J Infect Dis, 2004 Jan 15, 189(2), 328 - 38 Epub 2003 Dec 30.
Capsular expression in Streptococcus pneumoniae negatively affects spontaneous and antibiotic-induced lysis and contributes to antibiotic tolerance; Fernebro J et al.; Penicillin and vancomycin induce a lytic response in Streptococcus pneumoniae that requires the N-acetylmuramyl-l-alanine amidase LytA . We show that clinical isolates of pneumococci of capsular serotypes 1, 4, 6B, and 23F were generally less lytic to penicillin than pneumococci of serotypes 14 and 3 . In addition, most 9V isolates were less lytic to vancomycin, compared with isolates of other serotypes . Parent-mutant pairs expressing and not expressing capsular serotypes 2, 4, and 9V were compared for antibiotic-induced lysis . The nonencapsulated variants were considerably more lytic after beta-lactam and/or vancomycin treatment, and antibiotic tolerance was seen only in the context of capsule expression . Conversion from a nonlytic to a lytic phenotype, after loss of capsule expression, required an intact lytA autolysin gene . Exogenous addition of purified LytA gave a lower lytic response in capsulated strains, compared with that in nonencapsulated mutants . Spontaneous autolysis in stationary phase also was negatively affected by capsule expression in an autolysin-dependent manner . Long-term starvation in the stationary phase of the vancomycin- and penicillin-tolerant isolate I95 yielded nonencapsulated mutants that had lost antibiotic tolerance and were lytic to penicillin and vancomycin . The 9V capsular locus of I95 and one of these stationary phase-selected mutants were completely sequenced . The only difference found was a 1-bp frameshift deletion in the cps9vE gene of the lytic mutant, encoding a uridine diphosphate-glucosyl-1-phosphate transferase . Two additional independently isolated lytic mutants of I95 from the stationary phase also contained mutations in the same region of cps9vE, which identified it as a mutational hot spot . This report demonstrates that capsular polysaccharides negatively influence the lytic process and contribute to antibiotic tolerance in clinical isolates of pneumococci.

J Pediatr, 2004 Jan, 144(1), 68 - 74
Reduction in pediatric hospitalizations for varicella-related invasive group A streptococcal infections in the varicella vaccine era; Patel RA et al.; OBJECTIVES: To assess changes in hospitalization rates for invasive group A streptococcal (IGAS) and varicella-associated IGAS (VA-IGAS) infections at a pediatric hospital over a period of 9 years, to characterize clinical features of patients with IGAS infections, and to assess frequency of macrolide-resistant IGAS isolates.Study design Medical records of all hospitalized patients with group A streptococcus isolated from a normally sterile site from 1993 to 2001 were reviewed . Data collected included demographics, clinical course, microbiologic features, outcome, and presence of streptococcal toxic shock syndrome (STSS) or necrotizing fasciitis (NF) . Annual hospitalization rates for IGAS were determined . RESULTS: There were 144 patients with IGAS infections, including 11 (8%) with STSS or NF . Overall mortality rate was 2% (3/144) but 18% (2/11) among patients with STSS or NF . Preexisting varicella was present in 16% (23/144); 4 of 23 VA-IGAS cases had STSS or NF . Although there was no change in annual hospitalization rates for IGAS infections during the study period, the percentage of VA-IGAS hospitalizations decreased from 27% in the prevaccine era (1993 to 1995), to 16% during vaccine implementation (1996 to 1998) and 2% during widespread vaccine use (1999 to 2001) (linear-by-linear association, P=.001) . Macrolide resistance was low in 1993 to 1995 (5%, 1/19) and 1996 to 1998 (0%, 0/42) among tested IGAS isolates and increased significantly in 1999 to 2001 (13%, 5/38) (Fisher exact, P=.035) . CONCLUSIONS: A decline in pediatric varicella-related IGAS hospitalizations was temporally associated with utilization of varicella vaccine . These data reinforce the importance of universal varicella vaccination for children . Increasing macrolide resistance among IGAS isolates indicates a need for continued surveillance.

Emerg Infect Dis, 2003 Dec, 9(12), 1543 - 7
Comparative molecular and microbiologic diagnosis of bacterial endocarditis; Podglajen I et al.; Sequencing of 16S rDNA, and of sodAint and rpoBint in some cases, was applied to DNA from heart valves of 46 patients (36 with definite and 10 with possible endocarditis) . Sequence-based identifications were compared with those obtained with conventional methods . Among the 36 definite cases, 30 had positive blood cultures and 6 had negative cultures . Among the 30 positive cases, sequencing of 16S rDNA permitted identification of species (18), genus (8), or neither (4); sodAint and rpoBint sequencing was necessary for species identification in 8 cases . Species identifications were identical in only 61.5%, when conventional techniques and DNA sequencing were used . In five of the six blood culture-negative endocarditis cases, sequencing identified Bartonella quintana (3), B . henselae (1), and Streptococcus gallolyticus (1) . Our results demonstrate a clear benefit of molecular identification, particularly in cases of blood culture-negative endocarditis and of possible endocarditis, to confirm or invalidate the diagnosis . Moreover, in 19.4% of the definite cases, the improvement in species identification by sequencing led to improved patient management.

Value Health, 2004 Jan-Feb, 7(1), 36 - 51
A pharmacoeconomic evaluation of seven-valent pneumococcal conjugate vaccine in Spain; Asensi F et al.; BACKGROUND: Streptococcus pneumoniae is a leading cause of illness in children . Seven-valent pneumococcal conjugate vaccine (PCV-7), recently approved in the United States, is the first vaccine to provide protective immunity against pneumococcal disease in children under the age of 2 . PCV-7 is nearly 100% effective in preventing invasive pneumococcal infections and has been shown to significantly decrease the incidence of pneumonia and otitis media . OBJECTIVE: The objective of this study was to evaluate the health outcomes, costs, and cost-effectiveness of vaccination with PCV-7, compared with no vaccination for children in Spain . METHODS: A health state model was used to determine the health and economic outcomes in vaccinated and unvaccinated groups among children less than 5 years old . This analysis was conducted for a 10-year time horizon, beginning with initial vaccinations . Information on the burden of pneumococcal disease, in terms of data on the incidence and seroprevalence of disease, was collected from published and unpublished records, supplemented, and verified by Spanish pediatric and infectious disease experts . The efficacy of PCV-7 was based on updated findings of the Kaiser Permanente Efficacy Study . A cost-of-illness estimate for each pneumococcal disease was determined using decision tree analysis that considered direct and indirect costs . A birth cohort analysis compared the expected cost of vaccinated populations to age-matched unvaccinated populations . RESULTS: Implementing a PCV-7 vaccine program in Spain in a birth cohort of 360000 is expected to save approximately 16 lives and 132000 cases of pneumococcal disease over 10 years, resulting in total savings estimated at Euros 81 million (ESP13.5 billion), of which Euros 43.5 million (ESP7.1 billion) are direct medical savings . At a vaccine cost up to Euros 56.87 per dose (ESP9,462, the total cost of vaccinating a birth cohort of 360000 will be offset by the total savings owing to reduced morbidity . CONCLUSIONS: Implementing a universal PCV-7 vaccination program in Spain will significantly decrease the mortality and morbidity associated with pneumococcal infections in young children . At an assumed cost of Euros 48.56 (ESP8080) per dose, PCV-7 vaccination of Spanish children under the age of 5, followed over a 10-year period, is cost saving from the societal perspective and cost-effective from the payer perspective at Euros 22500 per LYG (ESP3,734713), comparing favorably with other preventive programs in Spain.

Am J Respir Med, 2003, 2(5), 385 - 94
Management of community-acquired pneumonia: a focus on conversion from hospital to the ambulatory setting; Tan JS et al.; Patients with community-acquired pneumonia (CAP) are treated in hospital or in the ambulatory care setting depending on the severity of illness . Despite numerous guidelines proposed, there is no agreement on specific criteria for hospitalization other than the clinicians' experience . The purpose of this review is to discuss the importance of the appropriate choice and timely administration of antibacterial agents, either in the hospital or in the outpatient setting.Since a high proportion of CAP patients will not have an etiologic agent identified at the time of initiation of treatment, the choice of antibacterial therapy is usually empiric . Antibacterial agents with activity against pneumococci and atypical pathogens causing pneumonia are the preferred choices . Macrolides, doxycycline, or respiratory fluoroquinolones have been recommended by various guidelines committees in North America for the treatment of pneumonia in patients with or without underlying comorbidities . Because of the increasing resistance to beta-lactams as well other antibacterial agents such as macrolides, doxycycline, and sulfamethoxazole/trimethoprim (cotrimoxazole), it is important that clinicians are aware of local statistics on resistance to Streptococcus pneumoniae, as infection with this bacterium is associated with high rates of morbidity and mortality . More recently, fluoroquinolone resistance has been reported, but the percentage of pneumococcal strains resistant to this agent is relatively low compared with the other antibacterial agents.Switch (intravenous to oral) therapy is recommended for hospitalized patients with CAP to facilitate early discharge, which has been shown to improve patient satisfaction and reduce hospital costs . Early conversion to oral therapy has not been shown to be associated with increased complications or higher mortality . Following prompt intravenous therapy and stabilization, patients with CAP should be treated with oral therapy in the ambulatory setting.

Am J Respir Med, 2003, 2(6), 459 - 68
Pseudomonal infections in patients with COPD: epidemiology and management; Lieberman D et al.; COPD is a common disease with increasing prevalence . The chronic course of the disease is characterized by acute exacerbations that cause significant worsening of symptoms . Bacterial infections play a dominant role in approximately half of the episodes of acute exacerbations of COPD . The importance of pseudomonal infection in patients with acute exacerbations of COPD stems from its relatively high prevalence in specific subgroups of these patients, and particularly its unique therapeutic ramifications . The colonization rate of Pseudomonas aeruginosa in patients with COPD in a stable condition is low.A review of a large number of clinical series of unselected outpatients with acute exacerbations of COPD revealed that P . aeruginosa was isolated from the patients' sputum at an average rate of 4% . This rate increased significantly in COPD patients with advanced airflow obstruction, in whom the rate of sputum isolates of P . aeruginosa reached 8-13% of all episodes of acute exacerbations of COPD . However, the great majority of bacteria isolated in these patients were not P . aeruginosa, but the three classic bacteria Streptococcus pneumoniae, Hemophilus influenzae, and Moraxella catarrhalis . The subgroup of patients, with acute exacerbations of COPD, with the highest rate of P . aeruginosa infection, which approaches 18% of the episodes, is mechanically ventilated patients . However, even in this subgroup the great majority of bacteria isolated are the above-mentioned three classic pathogens.In light of these epidemiologic data and other important considerations, and in order to achieve optimal antibacterial coverage for the common infectious etiologies, empiric antibacterial therapy should be instituted as follows . Patients with acute exacerbations of COPD with advanced airflow obstruction (FEV(1) <50% of predicted under stable conditions) should receive once daily oral therapy with one of the newer fluoroquinolones, i.e . levofloxacin, moxifloxacin, gatifloxacin, or gemifloxacin for 5-10 days . Patients with severe acute exacerbations of COPD who are receiving mechanical ventilation should receive amikacin in addition to one of the intravenous preparations of the newer fluoroquinolones or monotherapy with cefepime, a carbapenem or piperacillin/tazobactam . In both subgroups it is recommended that sputum cultures be performed before initiation of therapy so that the results can guide further therapy.

J Biol Chem, 2004 Apr 2, 279(14), 13896 - 901 Epub 2004 Jan 12.
Helix stability confers salt resistance upon helical antimicrobial peptides; Park IY et al.; Salt sensitivity of antimicrobial peptides poses a major obstacle in their development as novel antibiotics . Here we report the use of helix-capping motifs to confer salt resistance upon helical antimicrobial peptides . The helical content of the template peptide {RLLR}(5) was almost completely destroyed at salt concentrations over 200 mm NaCl, leading to a 8-32-fold decrease in antimicrobial activity . However, the introduction of helix-capping motifs at the helix termini resulted in a structurally stable peptide, which retained membrane-permeabilizing and antimicrobial activities upon exposure to salt . Furthermore, the peptide with helix-capping motifs directly inhibited the in vivo growth of Streptococcus pyogenes, which causes localized fasciitis in mice, and prevented the necrosis of the epidermis, dermis, and subcutaneous muscle layers . Results indicate that the adoption of helix-capping motifs into salt-sensitive antimicrobial peptides provides the necessary structural stability for the peptides to permeabilize cell membranes and cause cell death at physiological salt concentrations.

J Biol Chem, 2004 Mar 26, 279(13), 12462 - 8 Epub 2004 Jan 12.
Properties of GDP-mannose pyrophosphorylase, a critical enzyme and drug target in Leishmania mexicana; Davis AJ et al.; Leishmania parasites synthesize a range of mannose-containing glycoconjugates thought to be essential for virulence in the mammalian host and sandfly vector . A prerequisite for the synthesis of these molecules is the availability of the activated mannose donor, GDP-Man, the product of the catalysis of mannose-1-phosphate and GTP by GDP-mannose pyrophosphorylase (GDP-MP) . In contrast to the lethal phenotype in fungi, the deletion of the gene in Leishmania mexicana did not affect parasite viability but led to a total loss of virulence, making GDP-MP an ideal target for anti-Leishmania drug development . We show by immunofluorescence and subcellular fractionation that GDP-MP is a cytoplasmic protein, and we describe a colorimetric activity assay suitable for the high throughput screening of small molecule inhibitors . We expressed recombinant GDP-MP as a fusion with maltose-binding protein and separated the enzyme from maltose-binding protein by thrombin cleavage, ion-exchange, and size exclusion chromatography . Size exclusion chromatography and analytical ultracentrifugation studies demonstrate that GDP-MP self-associates to form an enzymatically active and stable hexamer . However, sedimentation studies show that the GDP-MP hexamer dissociates to trimers and monomers in a time-dependent manner, at low protein concentrations, at low ionic strength, and at alkaline pH . Circular dichroism spectroscopy reveals that GDP-MP is comprised of mixed alpha/beta structure, similar to its closest related homologue, N-acetyl-glucoseamine-1-phosphate uridyltransferase (Glmu) from Streptococcus pneumoniae . Our studies provide insight into the structure of a novel target for the development of anti-Leishmania drugs.

An Pediatr (Barc), 2004 Jan, 60(1), 75 - 9
{Group B Streptococcus late-onset disease presenting as cellulitis-adenitis syndrome}; Soler Palacin P et al.; Cellulitis-adenitis syndrome is a rare clinical manifestation of group B Streptococcus (GBS) late-onset disease . Its significance lies in the fact that local infection may be the only initial sign of systemic infection that is often concurrent with meningitis . Soft tissue involvement (cellulitis-adenitis) can sometimes be the only initial manifestation of GBS infection . We report four cases of GBS cellulitis-adenitis syndrome from different hospitals in Barcelona and Tarragona . We emphasize that early diagnosis and treatment may improve the potentially poor prognosis of these patients, and stress the need to rule out central nervous system involvement by studying cerebrospinal fluid.

J Paediatr Child Health, 2004 Jan-Feb, 40(1-2), 69 - 71
Fatal necrotizing pneumonia caused by group A streptococcus; Cengiz AB et al.; Group A streptococcus (GAS) causes invasive, non-invasive and non-suppurative diseases . Pneumonia is one of the invasive infections caused by GAS . Although GAS is a significant and serious cause of childhood pneumonia, it is often overlooked clinically . Similarly, the recent literature is surprisingly scant on reports of GAS pneumonia and concentrates mainly on varicella-associated invasive GAS diseases . In this case report, we present a previously healthy 7-year-old child with community-acquired pneumonia that progressed rapidly and resulted in sepsis, respiratory failure and death . In both blood and pleural fluid cultures, Streptococcus pyogenes were isolated . On autopsy, macroscopic examination revealed that the lung tissue appeared to have lost its normal architecture . Necrosis was present and the lung had a spongy appearance with some solid areas . The light microscopy revealed massive oedema, haemorrhages, intense inflammatory cell infiltration and necrosis . This case report highlights the need for consideration of invasive GAS infection in the event of severe, rapidly progressing pneumonia in children.

Drugs, 2004, 64(2), 159 - 204
Efflux-mediated drug resistance in bacteria; Li XZ et al.; Drug resistance in bacteria, and especially resistance to multiple antibacterials, has attracted much attention in recent years . In addition to the well known mechanisms, such as inactivation of drugs and alteration of targets, active efflux is now known to play a major role in the resistance of many species to antibacterials . Drug-specific efflux (e.g . that of tetracycline) has been recognised as the major mechanism of resistance to this drug in Gram-negative bacteria . In addition, we now recognise that multidrug efflux pumps are becoming increasingly important . Such pumps play major roles in the antiseptic resistance of Staphylococcus aureus, and fluoroquinolone resistance of S . aureus and Streptococcus pneumoniae . Multidrug pumps, often with very wide substrate specificity, are not only essential for the intrinsic resistance of many Gram-negative bacteria but also produce elevated levels of resistance when overexpressed . Paradoxically, 'advanced' agents for which resistance is unlikely to be caused by traditional mechanisms, such as fluoroquinolones and beta-lactams of the latest generations, are likely to select for overproduction mutants of these pumps and make the bacteria resistant in one step to practically all classes of antibacterial agents . Such overproduction mutants are also selected for by the use of antiseptics and biocides, increasingly incorporated into consumer products, and this is also of major concern . We can consider efflux pumps as potentially effective antibacterial targets . Inhibition of efflux pumps by an efflux pump inhibitor would restore the activity of an agent subject to efflux . An alternative approach is to develop antibacterials that would bypass the action of efflux pumps.

J Clin Microbiol, 2004 Jan, 42(1), 250 - 6
Serotype 14 variants of the France 9V(-3) clone from Baltimore, Maryland, can be differentiated by the cpsB gene; McEllistrem MC et al.; European serotype 14 variants of the France 9V(-3) clone, which have arisen through recombination events involving the penicillin binding protein 1a (pbp1a) gene, have cpsB sequences distinct from those of the 9V(-3) clone . Serotype 14 variants of the 9V(-3) clone have not been compared to genetically diverse serotype 14 strains isolated from an entire metropolitan area in the United States . All serotype 14 non-penicillin-susceptible Streptococcus pneumoniae strains causing invasive disease in Baltimore, Md., from 1995 to 1996 were compared by using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), pbp1a PCR restriction profiles, and cpsB and pbp1a sequences . The cpsB genes from strains of 13 serotypes also were analyzed to assess the correlation with serotype . Twenty-seven percent (3 of 11) of the serotype 14 strains were related by PFGE and MLST to the 9V(-3) clone . The serotype 14 variants from Baltimore, unlike the European variants, were related neither to the 9V(-3) clone nor to the R6 strain from positions 1498 to 1710 of the pbp1a gene . All serotype 14 strains had cpsB sequences that differed by <or=1% (0 to 5 of 476 bp) from each other and that were >or=16% (78 to 83 of 476 bp) divergent from that of the 9V(-3) clone . Allowing for a 2-bp difference in the cpsB sequence resulted in the highest correlation between the cpsB gene and serotype . Overall, 95% (84 of 88) of the strains were classified correctly by serotype with the cpsB sequence . The distal recombination site of the Baltimore serotype 14 variants of the 9V(-3) clone was not identical to that of the European serotype 14 variants . The cpsB gene was serotype specific regardless of whether capsular switching occurred . Although the correlation between serotype and the cpsB sequence was high, the overall diversity of the cpsB gene within a serotype likely will limit the role of this gene in a sequence-based serotyping method.

J Clin Microbiol, 2004 Jan, 42(1), 198 - 202
Demonstration of Streptococcus mutans with a cell wall polysaccharide specific to a new serotype, k, in the human oral cavity; Nakano K et al.; Streptococcus mutans organisms are occasionally isolated from the blood of patients with infective endocarditis, though the mechanisms of invasion and survival remain to be elucidated . Two of four blood isolates from patients with bacteremia or infective endocarditis (strains TW295 and TW871) were serologically untypeable by immunodiffusion testing, which was due to a lack of the glucose side chain of the serotype-specific polysaccharide antigen of S . mutans . Immunodiffusion analyses using antisera against these strains demonstrated that 2 of 100 isolates from 100 subjects showed a positive reaction, while further analysis of 2500 isolates from 50 subjects revealed that all 50 isolates from a single subject were not reactive with anti-c, -e, and -f antisera, though they were reactive with anti-TW295 and -TW871 antisera . The oral isolates showed biological properties similar to those of the reference S . mutans strain MT8148, including high levels of sucrose-dependent adhesion and cellular hydrophobicity, along with expression of glucosyltransferases and a protein antigen, PA . We designated these organisms serotype k . A glucose side chain-defective mutant strain was then constructed by insertional inactivation of the gluA gene of strain MT8148, which showed biological properties similar to those of serotype k of S . mutans . Serotype k oral isolates were less susceptible to phagocytosis, as were the gluA-inactivated mutant of strain MT8148 and blood isolates . These results indicate that S . mutans serotype k strains are present in the oral cavity in humans and may be able to survive longer in blood owing to their low susceptibility to phagocytosis.

Pediatr Nephrol, 2004 Mar, 19(3), 317 - 21 Epub 2004 Jan 09.
Pneumococcus-induced T-antigen activation in hemolytic uremic syndrome and anemia; Cochran JB et al.; The hemolytic uremic syndrome (HUS) is most commonly associated with Escherichia coli, but has been associated with other infections such as Streptococcus pneumoniae . Pneumococcus-induced HUS carries an increased risk of mortality and renal morbidity compared with E . coli-induced HUS . The pneumococcal organism produces an enzyme, which can expose an antigen (T-antigen) present on erythrocytes, platelets, and glomeruli . Antibodies to the T-antigen, normally found in human serum, bind the exposed T-antigen, and the resultant antigen-antibody reaction (T-activation) can lead to HUS and anemia . Clinicians need to be aware to request specific testing when pneumococcus-induced HUS/anemia is suspected, as current blood banking techniques do not routinely test for the presence of the T-antigen . Once this association is documented, washing all blood products and avoiding plasma products, if possible, is recommended . Plasmapheresis can be considered for the more critically ill patient . The incidence of pneumococcus-induced HUS may be increasing . We report six cases of pneumococcus-induced HUS/anemia presenting at our hospital.

Bull Exp Biol Med, 2003 Oct, 136(4), 423 - 6
Methodological approaches to deciphering the etiological structure of non-nosocomial pneumonia; Artemova LV et al.; Comparative analysis of the efficiency of PCR and microbiological methods for deciphering of the etiological structure of non-nosocomial pneumonias demonstrated the diagnostic significance of detecting Streptococcus pneumoniae DNA in the blood, but not sputum . Mechanisms of penetration of S . pneumoniae into the blood are discussed.

Ann Clin Microbiol Antimicrob . 2004 Jan 8;3(1):1.
Tracking the implementation of NCCLS M100-S12 expanded-spectrum cephalosporin MIC breakpoints for nonmeningeal isolates of Streptococcus pneumoniae by clinical laboratories in the United States during 2002 and 2003; Master RN et al.; BACKGROUND: The Performance Standards for Antimicrobial Susceptibility Testing, Twelfth Informational Supplement, M100-S12, published by the National Committee for Clinical Laboratory Standards (NCCLS) in January 2002 introduced distinct minimum inhibitory concentration (MIC) interpretative breakpoints for ceftriaxone, cefotaxime, and cefepime for nonmeningeal isolates of Streptococcus pneumoniae . Previously, a single set of interpretative breakpoints was used for both meningeal and nonmeningeal isolates . METHODS: To estimate the rate of adoption of the M100-S12 interpretive breakpoints by clinical laboratories, antimicrobial susceptibility test results for ceftriaxone and cefotaxime from nonmeningeal S . pneumoniae isolates were studied using data collected from January 2002 to June 2003 by The Surveillance Network(R) Database - USA (TSN(R)), an electronic surveillance database . RESULTS: Of the 262 laboratories that provided data that could be evaluated, 67.6% had adopted the M100-S12 breakpoints one and one-half years after they were published . CONCLUSIONS: The NCCLS M100-S12 recommendation to interpret MICs to expanded-spectrum cephalosporins using two distinct sets of breakpoints for meningeal and nonmeningeal isolates of S . pneumoniae was steadily implemented by clinical microbiology laboratories in the United States following their initial publication in January 2002 . The use of these new breakpoints more accurately reflects the clinical activities of expanded-spectrum cephalosporins than did the single set of interpretative breakpoints previously used for both meningeal and nonmeningeal isolates.

Protein Expr Purif, 2004 Feb, 33(2), 288 - 96
A GapC chimera retains the properties of the Streptococcus uberis wild-type GapC protein; Perez-Casal J et al.; The GapC products of Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis share considerable homology at the DNA and amino acid levels . The high similarity at the protein level suggests that one GapC protein might be used as a single antigen to protect dairy cows against infections with the contagious S . agalactiae and the environmental S . dysgalactiae and S . uberis strains . Despite their similarities, immunization with the S . dysgalactiae GapC did not protect dairy cows from a challenge with S . uberis, suggesting the presence of regions in GapC that are involved in species-specific protection . To produce a single antigen that can be used to protect against all streptococcal mastitis infections, we constructed a GapC chimeric protein using the S . uberis GapC product as the backbone followed by non-conserved peptide regions from the S . agalactiae and S . dysgalactiae GapC proteins . We report that the chimeric GapC protein retains the enzymatic activity of the S . uberis GapC protein . In addition, we fused the chimera to the OmpF and LipoF transport sequences of Escherichia coli and the GapC chimeras were present in membrane fractions of E . coli . These extracts could be the basis of an antigen preparation for use in mastitis vaccines.

Diagn Microbiol Infect Dis, 2003 Dec, 47(4), 587 - 93
In vitro pharmacodynamic activity of gatifloxacin, gemifloxacin, moxifloxacin and levofloxacin against Streptococcus pneumoniae containing specific mutations in DNA gyrase and topoisomerase IV; Garrison MW et al.; An in vitro pharmacodynatnic modeling apparatus (PDMA) generated specific bacterial kill profiles for single-dose regimens of gatifloxacin (GT), gemifloxacin (GM), moxifloxacin (MX) and levofloxacin (LV) against isolates of Streptococcus pneumoniae with specific QRDR profiles: SP-WT (no modifications); SP-C (changes in parC); and SP-AC (changes in both parC and gyrA) . No differences in 3-log reduction time or total log reduction were observed among the four agents for SP-WT; however, LV failed to achieve a 3-log reduction in SP-C and SP-AC, and total log reduction after 12 hrs was minimal compared to the other agents . GM and MX required less time for 3-log reduction of SP-AC compared to GT, but total log reductions in SP-AC were similar among the three newer quinolone agents (GM > MX > GT) . The study isolates with QRDR modifications greatly reduced LV activity . GM and MX maintained the greatest degree of activity against all study isolates and their activity was not adversely influenced by the genetic modifications in SP-C and SP-AC . The dual targeting characteristic of GM was also assessed, but did not offer significant advantages relative to MX and GT.

Diagn Microbiol Infect Dis, 2003 Dec, 47(4), 579 - 86
The activity of levofloxacin and other antimicrobials against clinical isolates of Streptococcus pneumoniae collected worldwide during 1999-2002; Jones ME et al.; Streptococcus pneumoniae is the most important causative bacterial pathogen in respiratory infections . Globally, increasing levels of resistant strains highlight the need for continued surveillance programs to guide antibiotic choice . The current study compared susceptibility results of 4,788 strains of S . pneumoniae collected during 2001-2002 to susceptibility results from 3,884 strains collected from the same hospitals during 1999-2000 . Participant centers were dispersed throughout five regions . By region, the prevalence of penicillin-resistant S . pneumoniae and percentage change from the previous 1999-2000 study was Mexico (26.0%, 12.5%), Brazil (7.9%; 5.5%), Asia (China, Hong Kong, South Korea, Thailand) (44.1%; 0.8%), Europe (France, Germany, Italy, Spain, UK) (11.1%; -0.6%) and South Africa (7.9; -1.8%) . Multidrug-resistant (MDR) strains of S . pneumoniae were most frequently isolated from Asia (36.3%) compared with approximately 5% in the other four regions . Increases in the incidence of MDR isolates in Mexico (13.5%), Brazil (1.7%) and Asia (6.1%) were reported with no increases in MDR in South Africa and Europe . Levofloxacin resistance was rarely associated with MDR phenotypes . Levofloxacin maintained an MIC(90) of 1 microg/ml against the isolates collected from all five regions with no change during the study periods, despite differences in levofloxacin resistance rates between regions or nations (0%-3.2%) . The prevalence of levofloxacin resistance (MIC > or =8 microg/ml) increased only slightly over the study period in Europe (0.3%-0.7%) and in Asia (3.0-3.2%), but little or no change was seen in Mexico (3.8%-0%) or Brazil or South Africa, where no levofloxacin resistant isolates were detected in either study period.

Commun Dis Public Health, 2003 Sep, 6(3), 221 - 7
Diagnosis of invasive pneumococcal infection by PCR amplification of Streptococcus pneumoniae genomic fragments in blood: a multi-centre comparative study; Sheppard CL et al.; The increasing interest in the prevention of pneumococcal disease by immunisation necessitates improved organism-specific surveillance . This is particularly the case with regard to the contribution of Streptococcus pneumoniae infection to community-acquired pneumonia where blood cultures are often negative and sputum culture results ambiguous . Examination by PCR of blood samples taken at hospital admission offers one possibility for such improvement . The sensitivity, specificity and convenience of three pneumolysin gene PCR assays were compared in a large study, using EDTA blood from 175 patients (95 with proven pneumococcal bacteraemia, 80 with bacteraemia due to other organisms) . The assays used were a PCR-enzyme immunoassay, a hybridisation probe assay run on the Roche LightCycler and a hydrolysis probe (TaqMan) assay run on an ABI 7700 . Overall samples from only 57% of patients with bacteraemic pneumococcal infection yielded a positive result in at least one assay . Individual sensitivities ranged from 45% (TaqMan/ABI) through 35% (PCR-EIA) to 21% (Hybridisation/LightCycler) . Specificity (PCR negative in the 80 control patients) ranged from 97-100% . The TaqMan/ABI assay was run in two centres and concordance between results was 91.4%, discrepancies being associated with very weakly positive samples . Overall, the TaqMan/ABI was the most sensitive and convenient assay; however, this method does not appear to offer any significant improvement over conventional blood cultures and is unlikely to be sufficiently sensitive to confirm a pneumococcal aetiology for non-bacteraemic pneumococcal pneumonia . For the present, therefore, blood culture is the preferred option.

Postgrad Med J, 2003 Dec, 79(938), 691 - 4
Pneumonia due to antibiotic resistant Streptococcus pneumoniae and Pseudomonas aeruginosa in the HAART era; Allen SH et al.; Antibiotic resistance profiles are useful in directing therapeutic strategies during bacterial infections . Patterns of antimicrobial resistance in Streptococcus pneumoniae and Pseudomonas aeruginosa associated pneumonia were investigated in an HIV-1 infected cohort during the era of highly active antiretroviral therapy . The median CD4 count at presentation was significantly lower for cases of P aeruginosa than for S pneumoniae . However, the number of antibiotic resistant cases of P aeruginosa decreased throughout the study period, while the incidence of S pneumoniae remained unchanged . In contrast to pneumococcal pneumonia, we show that antiretrovirals have protected from pneumonia due to antibiotic resistant P aeruginosa . These findings have implications for the treatment of individuals presenting with serious infections in which antibiotic therapy needs to be instituted before identification and sensitivities are known.

J Immunol, 2004 Jan 15, 172(2), 1198 - 202
L-ficolin specifically binds to lipoteichoic acid, a cell wall constituent of Gram-positive bacteria, and activates the lectin pathway of complement; Lynch NJ et al.; The lectin pathway of complement is activated when a carbohydrate recognition complex and associated serine proteases binds to the surface of a pathogen . Three recognition subcomponents have been shown to form active initiation complexes: mannan-binding lectin (MBL), L-ficolin, and H-ficolin . The importance of MBL in antimicrobial host defense is well recognized, but the role of the ficolins remains largely undefined . This report shows that L-ficolin specifically binds to lipoteichoic acid (LTA), a cell wall component found in all Gram-positive bacteria . Immobilized LTA from Staphylococcus aureus binds L-ficolin complexes from sera, and these complexes initiate lectin pathway-dependent C4 turnover . C4 activation correlates with serum L-ficolin concentration, but not with serum MBL levels . L-ficolin binding and corresponding levels of C4 turnover were observed on LTA purified from other clinically important bacteria, including Streptococcus pyogenes and Streptococcus agalactiae . None of the LTA preparations bound MBL, H-ficolin, or the classical pathway recognition molecule, C1q.

J Immunol, 2004 Jan 15, 172(2), 1125 - 31
The epidermal growth factor-seven transmembrane (EGF-TM7) receptor CD97 is required for neutrophil migration and host defense; Leemans JC et al.; The epidermal growth factor-seven transmembrane (EGF-TM7) family is a group of seven-span transmembrane receptors predominantly expressed by cells of the immune system . Family members CD97, EGF module-containing mucin-like receptor (EMR) 1, EMR2, EMR3, EMR4, and EGF-TM7-latrophilin-related protein are characterized by an extended extracellular region with a variable number of N-terminal EGF-like domains . EGF-TM7 receptors bind cellular ligands as demonstrated by the interaction of CD97 with decay accelerating factor (CD55) and dermatan sulfate . Investigating the effect of newly generated mAb on the migration of neutrophilic granulocytes, we here report for the first time in vivo data on the function of CD97 . In dextran sulfate sodium-induced experimental colitis, we show that homing of adoptively transferred neutrophils to the colon was significantly delayed when cells were preincubated with CD97 mAb . The consequences of this defect in neutrophil migration for host defense are demonstrated in a murine model of Streptococcus pneumoniae-induced pneumonia . Mice treated with CD97 mAb to EGF domain 1 (1B2) and EGF domain 3 (1C5) displayed a reduced granulocytic inflammatory infiltrate at 20 h after inoculation . This was associated with a significantly enhanced outgrowth of bacteria in the lungs at 44 h and a strongly diminished survival . Together, these findings indicate an essential role for CD97 in the migration of neutrophils.

Biol Psychiatry, 2004 Jan 1, 55(1), 61 - 8
Detecting pediatric autoimmune neuropsychiatric disorders associated with streptococcus in children with obsessive-compulsive disorder and tics; Murphy TK et al.; BACKGROUND: A subgroup of children with obsessive-compulsive and tic disorders are proposed to have an infectious trigger . The purpose of this study was to investigate the relationship between group A streptococcal titers and symptom fluctuations in children with a clinical course resembling that described for pediatric autoimmune neuropsychiatric disorders associated with streptococcus . METHODS: Twenty-five children with obsessive-compulsive disorder and/or tic disorder were evaluated for neuropsychiatric severity and group A streptococcal antibody titers (streptolysin O, deoxyribonuclease B, and carbohydrate A) at 6-week intervals for > or = six consecutive evaluations (total visits=277) . RESULTS: Children with large symptom fluctuations (n=15) were compared with children without dramatic fluctuations (n=10) . Co-movements of obsessive-compulsive/tic severity and group A streptococcal antibodies were assessed . In subjects with large symptom changes, positive correlations were found between streptococcal titers and obsessive-compulsive severity rating changes (p=.0130) . These subjects were also more likely to have elevated group A streptococcal titers during the majority of observations (p=.001) . Tic symptom exacerbations occurred more often in the fall/winter months than spring/summer months (p=.03) . CONCLUSIONS: Patients with marked obsessive-compulsive/tic symptom changes may be characterized by streptococcal titer elevations and exhibit evidence of seasonal tic exacerbations.

J Indian Soc Pedod Prev Dent, 2003 Sep, 21(3), 108 - 12
Microbial contamination of tooth brushes and their decontamination; Bhat SS et al.; With the dawn of the new century, dentistry has seen a new face in the fields of diagnosis, treatment and prevention . Twenty one children were asked to brush their teeth for five days . The brushes were put in Robertson's Cooked Meat broth and cultured . Growth of Streptococcus Mutans were seen . These brushes were then placed in disinfectants such as 0.2% chlorhexidine gluconate (Gp I), 1% sodium hypochlorite(Gp II) and water (Gp III) for 24 hrs and then cultured . Disappearance of growth of microorganisms was seen in Gp I and Gp II and remnants of growth seen in Gp III . It can be concluded that the use of disinfectant is a must for every individual at regular intervals.

Microbiology, 2004 Jan, 150(Pt 1), 189 - 98
The csp operon of Streptococcus salivarius encodes two predicted cell-surface proteins, one of which, CspB, is associated with the fimbriae; Levesque C et al.; A Tn917 mutant library was generated to identify genes involved in the biogenesis of Streptococcus salivarius fimbriae . A fimbria-deficient mutant was isolated by negative selection using an immunomagnetic separation technique with specific anti-fimbriae polyclonal antibodies (pAbs) . The transposon was inserted in an ORF, called orf176, which encoded a protein of unknown function . The transposon prevented the transcription of orf176 as well as two genes located downstream, which are designated cspA and cspB and which form the csp operon . Sequence analyses of CspA and CspB revealed that both proteins possessed the classic cell-wall-anchoring motif (LPXTG) of Gram-positive bacterial surface proteins . Recombinant CspA (rCspA) and CspB (rCspB) proteins were generated in Escherichia coli and used to produce pAbs . Immunolocalization experiments showed that anti-rCspB, but not anti-rCspA antibodies specifically recognized S . salivarius fimbriae . Our results suggested that the csp operon encoded predicted cell-surface proteins, one of which, CspB, was associated with the fimbriae.

Microbiology, 2004 Jan, 150(Pt 1), 103 - 8
Bovicin HJ50, a novel lantibiotic produced by Streptococcus bovis HJ50; Xiao H et al.; A bacteriocin-producing strain was isolated from raw milk and named Streptococcus bovis HJ50 . Like most bacteriocins produced by lactic acid bacteria, bovicin HJ50 showed a narrow range of inhibiting activity . It was sensitive to trypsin, subtilisin and proteinase K . Bovicin HJ50 was extracted by n-propanol and purified by SP Sepharose Fast Flow, followed by Phenyl Superose and Sephadex G-50 . Treatment of Micrococcus flavus NCIB8166 with bovicin HJ50 revealed potassium efflux from inside the cell in a concentration-dependent manner . The molecular mass of bovicin HJ50 was determined to be 3428.3 Da . MS analysis of DTT-treated bovicin HJ50 suggested that bovicin HJ50 contains a disulfide bridge . The structural gene of bovicin HJ50 was cloned by nested PCR based on its N-terminal amino acid sequence . Sequence analysis showed that it encodes a 58 aa prepeptide consisting of an N-terminal leader sequence of 25 aa and a C-terminal propeptide domain of 33 aa . Bovicin HJ50 shows similarity to type AII lantibiotics . Chemical modification using an ethanethiol-containing reaction mixture showed that two Thr residues are modified.

J Bacteriol, 2004 Jan, 186(2), 419 - 26
Identification of NAD+ synthetase from Streptococcus sobrinus as a B-cell-stimulatory protein; Veiga-Malta I et al.; Streptococcus sobrinus, one agent of dental caries, secretes a protein that induces lymphocyte polyclonal activation of the host as a mechanism of immune evasion . We have isolated from culture supernatants of this bacterium a protein with murine B-cell-stimulatory properties and subsequently cloned the relevant gene . It contains an open reading frame of 825 bp encoding a polypeptide with 275 amino acid residues and a molecular mass of 30 kDa . The protein displays high sequence homology with NAD(+) synthetases from several organisms, including a conserved fingerprint sequence (SGGXD) characteristic of ATP pyrophosphatases . The polypeptide was expressed in Escherichia coli as a hexahistidine-tagged protein and purified in an enzymatically active form . The recombinant NAD(+) synthetase stimulates murine B cells after in vitro treatment of spleen cell cultures, as demonstrated by its ability to induce up-regulation of the expression of CD69, an early marker of lymphocyte activation . Stimulation with the recombinant NAD(+) synthetase was also observed with other B-cell markers, such as CD19(+), B220(+), and CD21(+) . Cell proliferation follows the activation induced by the recombinant NAD(+) synthetase.

J Infect Dis, 2004 Jan 1, 189(1), 79 - 89 Epub 2003 Dec 22.
Identification of new candidate vaccine antigens made by Streptococcus pyogenes: purification and characterization of 16 putative extracellular lipoproteins; Lei B et al.; Putative extracellular lipoproteins made by group A Streptococcus (GAS) are the focus of this study, which was designed to identify new candidate vaccine antigens . Bioinformatic analysis of a serotype M1 GAS strain identified 30 open-reading frames encoding putative lipoproteins . The genes encoding the mature form of 29 of these proteins were cloned, and 16 recombinant proteins were overexpressed in Escherichia coli and purified to apparent homogeneity . The genes encoding these 16 proteins were highly conserved in GAS strains for which genome sequence data are available (serotypes M1, M3, M5, M12, M18, and M28) . Mice inoculated subcutaneously with GAS and humans with GAS pharyngitis and invasive infections seroconverted to most of the 16 recombinant proteins, which indicates that these lipoproteins were produced during infection . The blood of mice actively immunized with 5 of the 16 recombinant proteins had significantly (P<.05) increased growth-inhibitory activity, compared with the blood of unimmunized mice, which identified these proteins as potential new vaccine candidates.

J Biol Chem, 2004 Mar 12, 279(11), 9944 - 50 Epub 2003 Dec 29.
Mechanism of proton gating of a urea channel; Weeks DL et al.; The size and complexity of many pH-gated channels have frustrated the development of specific structural models . The small acid-activated six-membrane segment urea channel of Helicobacter hepaticus (HhUreI), homologous to the essential UreI of the gastric pathogen Helicobacter pylori, enables identification of all the periplasmic sites of proton gating by site-directed mutagenesis . Exposure to external acidity enhances {(14)C}urea uptake by Xenopus oocytes expressing HhUreI, with half-maximal activity (pH(0.5)) at pH 6.8 . A downward shift of pH(0.5) in single site mutants identified four of six protonatable periplasmic residues (His-50 at the boundary of the second transmembrane segment TM2, Glu-56 in the first periplasmic loop, Asp-59 at the boundary of TM3, and His-170 at the boundary of TM6) that affect proton gating . Asp-59 was the only site at which a protonatable residue appeared to be essential for pH gating . Mutation of Glu-110 or Glu-114 in PL2 did not affect the pH(0.5) of gating . A chimera, where the entire periplasmic domain of HhUreI was fused to the membrane domain of Streptococcus salivarius UreI (SsUreI), retained the pH-independent properties of SsUreI . Hence, proton gating of HhUreI likely depends upon the formation of hydrogen bonds by periplasmic residues that in turn produce conformational changes of the transmembrane domain . Further studies on HhUreI may facilitate understanding of other physiologically important pH-responsive channels.

Am J Ophthalmol, 2004 Jan, 137(1), 202 - 4
Endogenous endophthalmitis after otitis media; Siegersma JE et al.; PURPOSE: To report a case of bacterial endophthalmitis after otitis media in a healthy adult . DESIGN: Interventional case report . METHODS: A healthy 49-year-old Caucasian woman developed acute otitis media of the right ear . Three weeks after the first onset there was a recurrence of the otitis media, with perforation of the tympanic membrane . Two days after this, the woman presented at our clinic with endophthalmitis of the right eye . RESULTS: A culture of vitreous material grew Streptococcus pyogenes (Streptococcus Lancefield group A) . The same strain was found in a smear from the perforated ear . Despite aggressive treatment, the affected eye had to be eviscerated . CONCLUSIONS: Otitis media can result in a bacteremia . This may, even in a healthy adult, lead to a devastating endogenous bacterial endophthalmitis.

Am J Ophthalmol, 2004 Jan, 137(1), 43 - 51
Shifting trends in in vitro antibiotic susceptibilities for common ocular isolates during a period of 15 years; Chalita MR et al.; PURPOSE: To assess the in vitro susceptibility of the most common ocular bacterial isolates to several antibiotics and verify changing trends in the antibiotic susceptibility in a 15-year period . DESIGN: Experimental study . METHODS: All cultures positive for Staphylococcus aureus, coagulase-negative Staphylococcus (CNS), Streptococcus sp, and Pseudomonas sp in conjunctival (n = 4,585) and corneal (n = 3,779) samples from patients seen at the Federal University of Sao Paulo from 1985 to 2000 were evaluated . Cultures were performed in liquid and solid media, and susceptibility tests were done to amikacin, gentamicin, neomycin, tobramycin, ciprofloxacin, norfloxacin, ofloxacin, cephalothin, and chloramphenicol . RESULTS: Amikacin and neomycin showed an improvement of their sensitivity during the study period (88%-95% and 50%-85%, respectively) for corneal and conjunctival samples . Gentamicin and tobramycin revealed a decrease of sensitivity in time, from 95% to less than 80% in corneal and conjunctival samples . Ciprofloxacin, norfloxacin, and ofloxacin had good sensitivity to all evaluated bacteria, better in conjunctiva (95%) than in cornea (90%) . Sensitivity of S . aureus to cephalothin decreased during the study but was still 98% for CNS . Chloramphenicol had good sensitivity to S . aureus (85% in corneal and 92%in conjunctival samples), CNS (87% and 88.5%), and Streptococcus sp (95% and 96%) . CONCLUSIONS: Gentamicin, tobramycin, and cephalothin decreased their in vitro susceptibility to all tested pathogens . The fluoroquinolones remained a good choice in the treatment of ocular infections, with high suscep-tibility to all pathogens tested . Chloramphenicol also revealed an increase in its susceptibility to all bacteria evaluated.

Am J Ophthalmol, 2004 Jan, 137(1), 38 - 42
Endophthalmitis isolates and antibiotic sensitivities: a 6-year review of culture-proven cases; Benz MS et al.; PURPOSE: To investigate the spectrum of organisms causing culture-proven endophthalmitis and their sensitivities to commonly used antimicrobial agents . DESIGN: Retrospective, noncomparative, consecutive case series . METHODS: Medical records were reviewed of all patients with culture-proven endophthalmitis at a single institution between January 1, 1996, and December 31, 2001 . Endophthalmitis categories included postoperative, posttraumatic, endogenous, and miscellaneous (for example, keratitis) . The outcome measures included intravitreal isolates identified, antibiotic sensitivities, and category of endophthalmitis . RESULTS: In all, 313 organisms were isolated from 278 patients during the study interval . The most common organisms identified were Staphylococcus epidermidis in 27.8% (87/313), Streptococcus viridans group in 12.8% (40/313), other coagulase-negative staphylococci in 9.3% (29/313), Staphylococcus aureus in 7.7% (24/313), and Propionibacterium acnes in 7.0% (22/313) . Overall, 246 of 313 (78.5%) isolates were gram-positive organisms, 37 (11.8%) were gram-negative organisms, and 27 (8.6%) were fungi . For gram-positive organisms, sensitivities were the following: vancomycin 100%, gentamicin 78.4%, ciprofloxacin 68.3%, ceftazidime 63.6%, and cefazolin 66.8% . For gram-negative organisms, sensitivities were the following: ciprofloxacin 94.2%, amikacin 80.9%, ceftazidime 80.0%, and gentamicin 75.0% . Fungal isolates were Candida species (9/313), Aspergillus species (9/313), and other molds (9/313) . Among the endophthalmitis categories, the most frequent organisms were the following: (1) acute-onset postoperative: S epidermidis, 46.9%; (2) delayed-onset postoperative: S epidermidis, 22.7%; (3) delayed-onset bleb-associated: fastidious gram-negative rods, 20.4%; (4) posttraumatic: S epidermidis, 20.8%; (5) endogenous: Aspergillus species, 20.8%; and (6) miscellaneous: molds (other), 36.4% . CONCLUSIONS: In considering antibiotic treatment of endophthalmitis, it is important to recognize that no single antibiotic provided coverage for all of the microbes isolated from eyes with endophthalmitis . Combination therapy is recommended as the initial empiric treatment of suspected bacterial endophthalmitis . Appropriate history and characteristic clinical features may justify the use of initial antifungal agents . Knowledge of the most frequent causative organisms in various categories will help direct appropriate initial therapy.

J Coll Physicians Surg Pak, 2003 Nov, 13(11), 649 - 52
Treatment of necrotizing fasciitis with quinolones; Khan AT et al.; OBJECTIVE: To evaluate the effect of high dose quinolones therapy in patients with necrotizing fasciitis . DESIGN: Descriptive analytical study . PLACE AND DURATION OF STUDY: The department of Plastic and reconstructive surgery, Hayatabad Medical Complex, Peshawar, from January 2001 to March 2002 . SUBJECTS AND METHODS: Twenty consecutive patients, diagnosed with necrotizing fasciitis, were treated with intravenous quinolones (400 mg 8 hourly) . The response was evaluated in terms of subsidence of fever and C-reactive proteins levels . RESULTS: Majority of the patients was male (60%) . Lower limb involvement was most commonly involved (70%) . The most common initiating cause was injection abscess (45%) . Majority of the cultures showed polymicrobial infection (90%) . The most common isolate was Streptococcus pyogenes (65%) . Majority of the patients showed excellent response with intravenous quinolones (Ciprofloxacin) in high doses in 24-48 hours . Only two patients (10%) failed to respond to therapy due to severe infection and delay in seeking treatment . CONCLUSION: Intravenous quinolones (Ciprofloxacin) in high doses are effective in controlling necrotizing soft tissue infections.

Am Surg, 2003 Dec, 69(12), 1036 - 9; discussion 1039-40
Air following splenic embolization: infection or incidental finding?
Haan J, Bochicchio G, Kramer M, Scalea T.
The use of splenic embolization for nonoperative management has increased . With increased use of this adjunct, a new and frequent finding has been air within the areas of infarction in patients with or without clinical signs of infection . The purpose of this study was to determine if air within areas of splenic infarction is pathologic of infection or rather an incidental finding . A retrospective review over the past 3 years of inpatients undergoing splenic embolization and having pre- and postembolization abdominal computed tomography scans were reviewed for the findings of free air as well as any clinical signs of infection . A total of 96 consecutive patients were included . Of these, 12 had evidence of infarction with air . Six of these patients had undergone distal embolization with intraparenchymal air, but no symptoms . These were successfully observed . Two patients demonstrated subcapsular air/fluid levels, which underwent drainage with splenic preservation . Cultures were negative for infection . The remaining 4 underwent splenectomy . Of these, all had large collections of air . Two of these 4 spleens were infected: 1 with alpha-hemolytic Streptococcus and one with Clostridia perfringens . The remainder was sterile . This gave an overall infection rate of 17 per cent of patients with evidence of air . This yield increased to 33 per cent if the patient had symptoms and 50 per cent in those with large amounts of air and symptoms . Overall, we feel that air following embolization is a concern, but does not constitute infection . Patients with large amounts of air and signs and symptoms of infection will have a far higher infectious rate, 50 per cent in this limited series . In these patients, evaluation for infection is indicated; that being percutaneous sampling versus splenectomy.

Rev Pneumol Clin, 2003 Sep, 59(4), 209 - 12
{Streptococcus pneumoniae purulent pleurisy and hemolytic uremic syndrome . A case report}; Bouguila J et al.; A 22-month-old infant developed purulent pleurisy caused by Streptococcus pneumoniae and a hemolytic uremic syndrome . The diagnosis was suggested by the classical triad: hemolytic anemia, renal failure and thrombocytemia confirmed by renal biopsy which demonstrated extensive cortical necrosis . Renal involvement was severe, justifying an indication for renal transplantation.

Dev Comp Immunol, 2004 Apr, 28(4), 325 - 35
Characteristics and primary structure of a galectin in the skin mucus of the Japanese eel, Anguilla japonica; Tasumi S et al.; The characteristics and primary structure of AJL-1, one of the lectins in the skin mucus of the Japanese eel (Anguilla japonica), were examined . This lectin exhibited beta-galactoside specific activity in a Ca2+ independent manner . We previously reported that its molecular mass was 16,091Da, although it was approximately 30 kDa as determined by gel filtration, indicating that it is a homodimer having non-covalent bonds . This lectin was composed of 142 amino acid residues having no half-cystinyl residues, and showed homology to members of the galectin family, especially to proto-type galectins . Gene expression of this lectin was detected in skin only, and relative expression was high in an individual that exhibited resistance to infectious disease . AJL-1 showed agglutinating activity against pathogenic bacteria, Streptococcus difficile . This suggests that AJL-1 functions as an important defensive factor at the body surface.

Pediatr Crit Care Med, 2004 Jan, 5(1), 86 - 8
Cavernous sinus thrombosis complicating sinusitis; Cannon ML et al.; BACKGROUND: Septic cavernous sinus thrombosis is a rare complication of paranasal sinusitis . OBJECTIVE: To familiarize the clinician with the pathogenesis, diagnosis, and appropriate management of septic cavernous sinus thrombosis . DESIGN: Case report and literature review . SETTING: Pediatric intensive care unit in a university hospital . PATIENT: We present a 12-yr-old female with a 1 wk history of an upper respiratory tract infection with worsening dyspnea, cough, and swelling of the left eye progressing to adult respiratory distress syndrome . Secondary to the need for significant mechanical ventilatory support, venovenous extracorporeal membrane oxygenation was initiated . Computed tomography scan of the head and neck with contrast revealed bilateral cavernous sinus thrombosis . After broad-spectrum intravenous antibiotics and aggressive supportive care in conjunction with surgical intervention (maxillary sinus lavage and right orbital exploration) and anticoagulation therapy, the patient recovered . Blood cultures were positive for Viridans streptococcus . At discharge 3 wks later, the patient had improved, but had right-eye blindness . CONCLUSIONS: The diagnosis of septic cavernous sinus thrombosis requires a high index of suspicion and confirmation by imaging; early diagnosis and surgical drainage of the underlying primary source of infection in conjunction with long-term intravenous antibiotic therapy are critical for an optimal clinical outcome.

Am J Pathol, 2004 Jan, 164(1), 65 - 72
Antigen transport into Peyer's patches: increased uptake by constant numbers of M cells; Gebert A et al.; Membranous (M) cells are specialized epithelial cells of the Peyer's patches that sample antigens from the gut lumen, thereby enabling the host to respond immunologically . Recent studies suggest that this transport can be up-regulated within hours by de novo formation of M cells from enterocytes . To test this hypothesis, we used an in vivo model and induced the transcytosis of tracers in Peyer's patches by application of Streptococcus pneumoniae R36a into the gut lumen . Using cell-type-specific markers, we quantified M cells in the Peyer's patch domes, lymphocytes associated with M cells, and the transport rate for experimentally applied microbeads after 3 hours of exposure to R36a . The transport of latex microbeads was significantly increased by +131% in the R36a-treated patches as compared to buffer controls (P < 0.001) . While in controls, each M cell was associated with 2.05 +/- 0.64 lymphocytes, a significant increase (+55.1%; P < 0.001) was determined in the R36a-treated patches . However, no statistical difference was detected in the percentage of M cells in the dome epithelia (46.0 +/- 4.6% versus 45.5 +/- 3.8%) . It is concluded that bacteria-induced up-regulation of particle transport in Peyer's patch domes is due to an increased transport rate of the M cells, but not to a de novo formation of M cells . The data support the hypothesis that M cells represent a separate cell lineage that does not derive from enterocytes on the domes.

Scand J Prim Health Care, 2003 Dec, 21(4), 196 - 8
Reduction in antibiotic prescribing for respiratory tract infections is needed!
Molstad S.
In general practice, approximately 25% of consultations are related to infectious diseases . The emergence and spread of resistant bacteria are related to antibiotic use on an individual and on a community level . Antibiotic prescribing differs profoundly from one European country to the next, though there is no evidence of differences in the prevalence of infectious diseases . Most respiratory tract infections are self-limiting conditions, and recent evidence shows that antibiotics only slightly modify the course of most respiratory tract infections . The general practitioner should focus on patients with more severe symptoms who might benefit more from antibiotic treatment . In general, antibiotics should be prescribed for acute pneumonia . In addition, we may offer antibiotics to a selected group of patients with more severe symptoms of maxillary sinusitis, pharyngotonsillitis and acute exacerbation of chronic obstructive pulmonary disease/chronic bronchitis . In the diagnostic procedure, rapid tests of Streptococcus pyogenus and C-reactive protein may be valuable in carefully selected cases . Penicillins (penicillin V, amoxycillin) should be the first choice in most respiratory tract infections . Larger studies in general practice are needed to analyse the impact of antibiotic prescribing on morbidity, the occurrence of rare complications and spread of resistance . The greatest challenge will be to implement current knowledge in daily praxis.

Pharmacotherapy, 2003 Dec, 23(12), 1531 - 7
Linezolid, levofloxacin, and vancomycin against vancomycin-tolerant and fluoroquinolone-resistant Streptococcus pneumoniae in an in vitro pharmacodynamic model; Cha R et al.; STUDY OBJECTIVE: To compare the pharmacodynamic profiles of linezolid, levofloxacin, and vancomycin against clinical strains of Streptococcus pneumoniae, including vancomycin-tolerant and fluoroquinolone-resistant isolates . DESIGN: In vitro pharmacodynamic model . SETTING: Biosafety level 2, university research laboratory . BACTERIAL STRAINS: Ciprofloxacin-susceptible (79), ciprofloxacin-resistant (R921), and vancomycin-tolerant (P9802-020) clinical strains of S . pneumoniae . INTERVENTION: An in vitro pharmacodynamic model was used to simulate standard dosing regimens of linezolid, levofloxacin, and vancomycin against the isolates 79, R921, and P9802-020 . MEASUREMENTS AND MAIN RESULTS: Bacterial density was profiled over 48 hours . Minimum inhibitory concentrations (MICs) for linezolid, levofloxacin, and vancomycin, respectively were 1, 1, 0.5 microg/ml for isolate 79; 1, 4, 0.5 microg/ml for R921; and 0.5, 0.5, 0.5 microg/ml for P9802-020 . Vancomycin minimum bactericidal concentration (MBC) values varied across large ranges for the tested strains . Linezolid achieved 99.9% kill against 79 and R921 by 24 and 28 hours, respectively . Levofloxacin achieved 99.9% kill against 79 and P9802-020 by 28 and 4 hours, respectively . Vancomycin achieved 99.9% kill against 79 and R921 by 8 and 24 hours, respectively . Levofloxacin did not demonstrate activity against R921 at the 48-hour end point . Minimal kill (< 2 log) at 48 hours was noted for vancomycin and linezolid against P9802-020 . Conclusion . Vancomycin tolerance appeared to be more reliably characterized by persistent viability in time-kill analyses than by MBC:MIC ratios . Vancomycin exhibited bactericidal activity against the non-vancomycin-tolerant strains of S . pneumoniae . Linezolid exhibited both bactericidal and bacteriostatic activity against all three strains tested, whereas levofloxacin demonstrated bactericidal activity against the fluoroquinolone-susceptible isolates . Further investigation of treatment alternatives for infections due to vancomycin-tolerant S . pneumoniae are needed.

J Matern Fetal Neonatal Med, 2003 Sep, 14(3), 151 - 7
Maternal and fetal inflammatory responses in unexplained fetal death; Blackwell S et al.; OBJECTIVE: The role of intra-amniotic infection in the etiology of fetal death has been proposed . This study was conducted to determine the prevalence of microbial invasion of the amniotic cavity (MIAC) and the frequency of maternal and/or fetal inflammation in patients presenting with a fetal death . METHODS: A prospective study was conducted in patients with a fetal death . Amniocenteses were performed for clinical indications (karyotype), as well as to assess the microbiological and cytological state of the amniotic cavity . Fluid was cultured for aerobic and anaerobic bacteria and genital mycoplasmas . An amniotic fluid white blood cell count and glucose determinations were also performed . Histological examination of the placenta was conducted to identify a maternal inflammatory response (acute chorioamnionitis) or a fetal inflammatory response (funisitis) . RESULTS: This study included 44 patients with intrauterine fetal death . The median gestational age at diagnosis was 30.1 weeks (range 16.3-40.4 weeks) . One patient had documented MIAC (1/44) . Acute histological chorioamnionitis was found in 20.9% (9/43), but a fetal inflammatory response was observed in only 2.3% (1/43) of cases . One patient had a positive amniotic fluid culture for Streptococcus agalactiae (group B streptococcus) . CONCLUSION: Histological chorioamnionitis was present in 20.9% of cases, but MIAC could be demonstrated with conventional microbiological techniques in only one case . A fetal inflammatory response was nine times less frequent than a maternal inflammatory response (maternal 20.9% vs . fetal 2.3%, p = 0.008) in cases of fetal death.

Proc Natl Acad Sci U S A, 2004 Jan 6, 101(1), 215 - 20 Epub 2003 Dec 23.
The C-type lectin SIGN-R1 mediates uptake of the capsular polysaccharide of Streptococcus pneumoniae in the marginal zone of mouse spleen; Kang YS et al.; SIGN-R1, a recently discovered C-type lectin expressed at high levels on macrophages within the marginal zone of the spleen, mediates the uptake of dextran polysaccharides by these phagocytes . We now find that encapsulated Streptococcus pneumoniae are rapidly cleared by these macrophages from the bloodstream, and that capture also takes place when different cell lines express SIGN-R1 after transfection . To assess the role of the capsular polysaccharide of S . pneumoniae (CPS) in the interaction of SIGN-R1 with pneumococci, we first studied binding and uptake of serotype 14 CPS in transfected cells . Binding was observed and was of a much higher avidity (3000-fold) for CPS 14 than dextran . The CPSs from four different serotypes were also cleared by marginal zone macrophages in vivo . To establish a role for SIGN-R1 in this uptake, we selectively down-regulated expression of the lectin by pretreatment of the mice with SIGN-R1 antibodies, including a newly generated hamster monoclonal called 22D1 . For several days after this transient knockout, the marginal zone macrophages were unable to take up either CPSs or dextrans . Therefore, marginal zone macrophages in mice have a receptor that interacts with capsular pneumococcal polysaccharides, setting the stage for further studies of the functional consequences of this interaction.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 80 - 5
Efficacy of BB-83698, a novel peptide deformylase inhibitor, in a mouse model of pneumococcal pneumonia; Azoulay-Dupuis E et al.; The efficacy of BB-83698, a novel potent peptide deformylase inhibitor, was evaluated in a mouse model of acute pneumonia . The Streptococcus pneumoniae isolates tested included four virulent strains (one penicillin-susceptible wild-type strain, one macrolide-resistant strain, and two quinolone-resistant mutants {a mutant carrying mutations in ParC and GyrA and an efflux mutant} isogenic to the wild type) and two poorly virulent penicillin-resistant strains . Pneumonia was induced by intratracheal inoculation of 10(5) CFU (virulent strains) into immunocompetent mice or 10(7) CFU (less virulent strains) into leukopenic mice . Animals received three or six subcutaneous injections of antibiotics at 12- or 24-h intervals, with antibiotic treatment initiated at 3, 6, 12, or 18 h postinfection (p.i.) . BB-83698 showed potent in vitro activity against all strains (MICs, 0.06 to 0.25 micro g/ml) . In the in vivo model, all control animals died within 2 to 5 days of infection . BB-83698 (80 mg/kg of body weight twice daily or 160 mg/kg once daily) protected 70 to 100% of the animals, as measured 10 days p.i., regardless of the preexisting resistance mechanisms . In contrast, the survival rates for animals treated with the comparator antibiotics were 30% for animals treated with erythromycin (100 mg/kg) and infected with the macrolide-resistant strain, 34% for animals treated with amoxicillin (200 mg/kg every 8 h) and infected with the penicillin-resistant strain, and 0 and 78% for animals treated with ciprofloxacin (250 mg/kg) and infected with the ParC and GyrA mutant and the efflux mutant, respectively . At 80 mg/kg, BB-83698 generated a peak concentration in lung tissue of 61.9 micro g/ml within 1 h and areas under the concentration-times curves of 57.4 and 229.4 micro g . h/ml for plasma and lung tissue, respectively . The emergence of S . pneumoniae isolates with reduced susceptibilities to BB-83698 was not observed following treatment with a suboptimal dosing regimen . In conclusion, the potent in vitro activity of BB-83698 against S . pneumoniae, including resistant strains, translates into good in vivo efficacy in a mouse pneumonia model.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 63 - 8
In vivo pharmacodynamic activity of daptomycin; Safdar N et al.; Daptomycin is a lipopeptide antibiotic with activity against a wide range of gram-positive bacteria . We used the neutropenic murine thigh model to characterize the pharmacodynamics of daptomycin . ICR/Swiss mice were rendered neutropenic with cyclophosphamide; and the thigh muscles of the mice were infected with strains of Staphylococcus aureus, Streptococcus pneumoniae, and Enterococcus faecium . Animals were treated by subcutaneous injection of daptomycin at doses of 0.20 to 400 mg/kg of body weight/day divided into one, two, four, or eight doses over 24 h . Daptomycin exhibited linear pharmacokinetics, with an area under the concentration-time curve (AUC) from time zero to infinity/dose of 9.4 and a half-life of 0.9 to 1.4 h . The level of protein binding was 90% . Free daptomycin exhibited concentration-dependent killing and produced in vivo postantibiotic effects (PAEs) of 4.8 to 10.8 h . Nonlinear regression analysis was used to determine which pharmacokinetic (PK) or pharmacodynamic (PD) parameter was important for efficacy by using free drug concentrations . The peak concentration/MIC (peak/MIC) ratio and 24-h AUC/MIC ratio were the PK and PD parameters that best correlated with in vivo efficacy (R(2) = 83 to 87% for peak/MIC and R(2) = 86% for the AUC/MIC ratio, whereas R(2) = 47 to 50% for the time that the concentration was greater than the MIC) against standard strains of S . aureus and S . pneumoniae . The peak/MIC ratios required for a bacteriostatic effect ranged from 12 to 36 for S . pneumoniae, 59 to 94 for S . aureus, and 0.14 to 0.25 for E . faecium . The AUC/MIC ratios needed for a bacteriostatic effect ranged from 75 to 237 for S . pneumoniae, 388 to 537 for S . aureus, and 0.94 to 1.67 for E . faecium . The free daptomycin concentrations needed to average from one to two times the MIC over 24 h to produce a bacteriostatic effect and two to four times the MIC over 24 h to produce greater than 99% killing . The long PAE and potent bactericidal activity make daptomycin an attractive option for the treatment of infections caused by gram-positive bacteria.

Pediatr Pathol Mol Med, 2003 Jul-Aug, 22(4), 303 - 9
Classification of M nontypeable group A streptococcus with the use of field inversion gel electrophoresis; Martin JM et al.; Group A streptococcus (GAS) are traditionally classified based on M and T protein antigens . However, many strains do not react with available M antisera and are classified as M nontypeable (M NT) . M and T typing, field inversion gel electrophoresis (FIGE), and 5' emm gene sequencing were performed on 24 M NT GAS isolates . FIGE patterns of the M NT isolates were compared by visual inspection and by computer analysis with the patterns of 139 isolates representing 72 M-types of GAS . Seven different FIGE patterns (I-VII) were seen among the M NT isolates . FIGE patterns I and III were identical or closely related to patterns seen with M12 and M22 isolates . The computer analysis determined the following relationships: pattern IV to M5, pattern VI to M61, and pattern VII to M59 . Thus, the emm gene sequence correlated with the computer analysis of the FIGE pattern for each isolate . FIGE can be useful to help distinguish clinical isolates of GAS in an epidemiological study.

Int J Infect Dis, 2004 Jan, 8(1), 53 - 8
Pneumococcal bacteremia in a single center in Argentina; Altclas J et al.; OBJECTIVE: To determine the clinical and microbiologic characteristics of pneumococcal bacteremia at Sanatorio Mitre, Buenos Aires, Argentina . METHODS: One-hundred-and-seven episodes of pneumococcal bacteremia were prospectively analyzed from 1993 to 1998 . Demographics, clinical and microbiological variables were studied . RESULTS: Eighty-one patients (76%) were adults and 26 children (24%) . Most cases (98%) were acquired in the community . Seventy-nine patients (74%) had at least one underlying condition . The primary source of bacteremia was found in 91 patients (85%), the lungs being the most common source . Streptococcus pneumoniae was isolated from one sterile site other than the primary focus in 25 patients (23%) . Eighty-five (79%) of the Streptococcus pneumoniae were susceptible to penicillin and 22 (21%) showed intermediate or high resistance to penicillin and 2% were additionally resistant to ceftriaxone.Initial antimicrobial therapy was appropriate in 95% of the cases . The overall mortality was 21%, however adults admitted to the intensive care unit (ICU) had higher mortality (81%) . No patients under 14 years old died . Multivariate analysis showed that age and recovery of the organisms from a sterile site other than the primary focus were statistically significant predictors of mortality . CONCLUSION: Bacteremic pneumococcal infections continue to be an important worldwide problem causing morbidity and high mortality despite supportive care and appropriate antimicrobial therapy.

J Pediatr (Rio J), 1995 May-Jun, 71(3), 166 - 8
{Neonatal arthritis of the hip due to group B streptococcus}; Campos JM et al.; The authors describe a case of neonatal osteoarthritis of the hip due to group B streptococcus and discuss the importance of recognizing this bacteria in the late focal infections of the newborn.

Pediatr Infect Dis J, 2003 Dec, 22(12), 1143 - 51
Appropriate antibiotic use and why it is important: the challenges of bacterial resistance; Lieberman JM; After the introduction of antibiotics in the mid-20th century, clinicians soon witnessed clinical failures secondary to bacterial resistance . Despite scientists' efforts to synthesize more potent antibiotics during the last five decades, bacterial resistance continues to evolve, in large part because of the overuse and misuse of antibiotics . The treatment of several pathogens, including methicillin-resistant Staphylococcus aureus, penicillin-resistant Streptococcus pneumoniae and vancomycin-resistant enterococci, is problematic . New solutions are needed to preserve the activity of our current antibiotic armamentarium, to lower the overall risk of bacterial resistance and to successfully treat patients with resistant bacterial infections . Options include: development of new antibiotics to treat resistant organisms; vaccination to prevent infections; and improved use of antibiotics . Because bacteria will eventually develop means to avoid being killed by antibiotics, judicious use of antibiotics by all clinicians is imperative . Appropriate antibiotic use involves selection of a "targeted spectrum" antibiotic, as well as an appropriate dose and duration.

Pediatr Infect Dis J, 2003 Dec, 22(12), 1069 - 74
Invasive pneumococcal disease in Costa Rican children: a seven year survey; Ulloa-Gutierrez R et al.; BACKGROUND: Streptococcus pneumoniae is a leading cause of invasive bacterial disease in children worldwide . Although morbidity and mortality associated with invasive pneumococcal disease (IPD) are known to be high in Latin American infants, the current situation for Central American children is unclear . METHODS: A 7-year retrospective review of IPD cases (January 1995 to December 2001) treated at the National Children's Hospital in San Jose, Costa Rica . RESULTS: We analyzed 135 episodes that occurred in 132 patients . The mean age of presentation was 35.7 months (range, 0 to 11.4 ys), with 73.3% of all episodes occurring in patients <5 years of age and 56% occurring in patients <24 months of age . Underlying medical conditions were present in 47% of children . The most common clinical presentations were meningitis in 56 (41.5%) patients, pneumonia in 36 (26.7%), bacteremia alone in 30 (22.2%), peritonitis in 10 (7.4%), septic arthritis in 2 (1.5%) and osteomyelitis in 1 (0.7%) . The case fatality rate was 14.4%, and children <2 years of age had the highest rates of complications, sequelae and death . Penicillin or cefotaxime nonsusceptibility was observed in 14.3% (10.7% intermediate, 3.6% resistant) and 4.5% (1.5% intermediate, 3% resistant) of tested isolates, respectively . CONCLUSIONS: IPD in Costa Rica is associated with high morbidity and mortality, particularly among young infants . Most prevalent IPD are the ones observed in developed countries before the introduction of current conjugated vaccine . Penicillin and third generation cephalosporin resistance in invasive cases is present at low rates.

J Immunol, 2004 Jan 1, 172(1), 532 - 9
Differential regulation of IgG anti-capsular polysaccharide and antiprotein responses to intact Streptococcus pneumoniae in the presence of cognate CD4+ T cell help; Khan AQ et al.; The relative lack of memory for IgG antipolysaccharide responses is believed to be secondary to the inability of polysaccharides to associate with MHC class II molecules and thus a failure to recruit cognate CD4+ T cell help . However, little is known concerning the role of T cells and the generation of memory for antipolysaccharide Ig responses to intact extracellular bacteria . We used heat-killed, intact Streptococcus pneumoniae, capsular type 14 (Pn14), to evaluate the IgM and IgG responses specific for the capsular polysaccharide (PPS14), the phosphorylcholine determinant of the cell wall C-polysaccharide, and the cell wall protein, pneumococcal surface protein A (PspA) . We demonstrate that the IgG (but not IgM), anti-PPS14, and anti-PspA responses to Pn14 are CD4+ T cell dependent and TCR specific . Nevertheless, in contrast to the anti-PspA response, the IgG anti-PPS14 response shows no apparent memory, an accelerated kinetics of primary Ig induction, and a more rapid delivery of CD4+ T cell help . In contrast, the IgG anti-phosphorylcholine response, although also dependent on CD4+ T cells, is TCR nonspecific . We make similar observations using soluble conjugates of PPS14-PspA and C-polysaccharide-PspA . These data lead us to suggest that the central issue concerning the mechanisms underlying different functional outcomes for anti-bacterial IgG responses to capsular polysaccharide vs protein Ags is not necessarily based on the ability to recruit cognate CD4+ T cell help, but perhaps on the nature of the B cell Ag receptor signaling that occurs and/or on the responding B cell subpopulations.

J Immunol, 2004 Jan 1, 172(1), 426 - 32
Organism-specific neutrophil-endothelial cell interactions in response to Escherichia coli, Streptococcus pneumoniae, and Staphylococcus aureus; Moreland JG et al.; The recruitment of polymorphonuclear leukocytes (PMNs) from the vascular space into the lung interstitium and airspace is an early step in the host innate immune response to bacterial invasion of these sites . To determine the ability of intact bacteria to directly elicit PMN migration across an endothelial monolayer, we studied in vitro migration of PMNs across a monolayer of human pulmonary microvascular endothelial cells in response to Streptococcus pneumoniae, Staphylococcus aureus, and Escherichia coli, as well as to purified E . coli LPS . Bacterial induction of PMN migration was dose dependent and elicited by > or =10(4) bacteria/ml of each of the species tested . Pretreatment of PMNs with blocking Abs to CD18 significantly inhibited migration of PMN in response to all stimuli tested, but had the most profound effect on migration to S . pneumoniae and S . aureus . Intact E . coli were 10 times more potent in inducing transmigration of PMNs than a corresponding amount of purified LPS . Bacterial induction of PMN migration did not correlate with up-regulation of surface endothelial ICAM-1 expression (purified LPS >> intact E . coli > S . aureus and S . pneumoniae) nor up-regulation of VCAM-1 and E-selectin . Neutralizing Ab to ICAM-1 had no effect on PMN migration to any of the bacteria or to purified LPS . These findings demonstrate that diverse bacterial pathogens induce PMN migration across a pulmonary microvascular endothelial cell monolayer in a fashion that appears to be organism specific . In addition, intact bacteria elicit PMN-endothelial cell interactions distinct from those seen when purified bacterial products are used as agonists.

Infect Immun, 2004 Jan, 72(1), 606 - 10
Acquisition of host plasmin activity by the Swine pathogen Streptococcus suis serotype 2; Jobin MC et al.; In this study, the plasminogen-binding activity of Streptococcus suis serotype 2 was investigated . Bound human plasminogen was activated by purified streptokinase, urokinase, or Streptococcus dysgalactiae subsp . equisimilis culture supernatant . Both human and porcine plasminogen were bound by S . suis . Binding was inhibited by epsilon-aminocaproic acid, and the plasminogen receptor was heat and sodium dodecyl sulfate resistant . One of the receptors was identified as glyceraldehyde-3-phosphate dehydrogenase . S . suis-associated plasmin activity was capable of activating free plasminogen, which in turn could contribute to degradation of fibronectin . This is the first report on the plasminogen-binding activity of S . suis . Further studies may reveal a contribution of this activity to the virulence of S . suis.

Infect Immun, 2004 Jan, 72(1), 408 - 13
Contribution of glutathione peroxidase to the virulence of Streptococcus pyogenes; Brenot A et al.; Glutathione peroxidases are widespread among eukaryotic organisms and function as a major defense against hydrogen peroxide and organic peroxides . However, glutathione peroxidases are not well studied among prokaryotic organisms and have not previously been shown to promote bacterial virulence . Recently, a gene with homology to glutathione peroxidase was shown to contribute to the antioxidant defenses of Streptococcus pyogenes (group A streptococcus) . Since this bacterium causes numerous suppurative diseases that require it to thrive in highly inflamed tissue, it was of interest to determine if glutathione peroxidase is important for virulence . In this study, we report that GpoA glutathione peroxidase is the major glutathione peroxidase in S . pyogenes and is essential for S . pyogenes pathogenesis in several murine models that mimic different aspects of streptococcal suppurative disease . In contrast, glutathione peroxidase is not essential for virulence in a zebrafish model of streptococcal myositis, a disease characterized by the absence of an inflammatory cell infiltrate . Taken together, these data suggest that S . pyogenes requires glutathione peroxidase to adapt to oxidative stress that accompanies an inflammatory response, and the data provide the first demonstration of a role for glutathione peroxidase in bacterial virulence . The fact that genes encoding putative glutathione peroxidases are found in the genomes of many pathogenic bacterial species suggests that glutathione peroxidase may have a general role in bacterial pathogenesis.

Infect Immun, 2004 Jan, 72(1), 352 - 8
Innate immune response of oral and foreskin keratinocytes: utilization of different signaling pathways by various bacterial species; Chung WO et al.; The innate immune response is critical for the epithelial antimicrobial barrier . The human beta-defensins are small, cationic antimicrobial peptides that are made by epithelial cells and that play a role in mucosal and skin defenses . Human beta-defensin 1 (hBD-1) is expressed constitutively in epithelial tissues, whereas hBD-2 and hBD-3 are expressed in response to bacterial stimuli or inflammation . Previous studies showed that hBD-2 was induced by Fusobacterium nucleatum cell wall extract without the involvement of the NF-kappaB transcription factors, which typically are associated with innate immunity and inflammation . The goal of this study was to characterize signaling pathways involved in hBD-2 induction in response to commensal and pathogenic bacteria . Cultured human oral and foreskin keratinocytes were treated separately with inhibitors of NF-kappaB, c-Jun N-terminal kinase (JNK), and p38 and then stimulated with oral commensal Streptococcus gordonii, oral pathogens Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, skin commensal Staphylococcus epidermidis, or skin pathogen Streptococcus pyogenes . Different bacteria induced different levels of hBD-2 and in response to the various inhibitors tested, although certain common patterns were observed for commensal- and pathogen-stimulated cells . hBD-2 induction by all bacteria tested was partially or completely blocked by inhibitors of the JNK and p38 pathways . However, in addition, hBD-2 induction by pathogenic bacteria in both oral and foreskin keratinocytes was blocked by inhibitors of NF-kappaB . The results indicate that commensal and pathogenic bacteria utilize different pathways in hBD-2 induction and suggest that epithelial cells from different body sites have common signaling mechanisms to distinguish between commensal and pathogenic bacteria.

Infect Immun, 2004 Jan, 72(1), 114 - 22
Effects of PspA and antibodies to PspA on activation and deposition of complement on the pneumococcal surface; Ren B et al.; Streptococcus pneumoniae infection is a frequent cause of pneumonia, otitis media, meningitis, and septicemia . Pneumococcal surface protein A (PspA) is an important virulence factor on the pathogen surface, and it is known to interfere with complement activation . In this study, flow cytometry was used to study the effects of PspA and antibodies to PspA on the deposition of complement C3 on the surface of a capsular type 3 strain, WU2, and its PspA- mutant, JY1119 . Using naive mouse serum as a complement source, measurable deposition of C3 was observed within 4 min on PspA- pneumococci, and the amount of surface-bound C3 accumulated rapidly as the amount of serum was increased . In contrast, very little C3 was deposited on the PspA+ strain . In nonimmune mouse serum, the classical pathway was the dominant activation pathway triggered by PspA- pneumococci . Accordingly, EGTA blocked almost all of the complement activation . Moreover, a significant amount of C3 was still deposited on the PspA- strain when serum from factor B-deficient mice was used . This deposition was not observed on the PspA+ pneumococci, indicating that PspA may inhibit complement deposition via the classical pathway . Furthermore, under the conditions we tested, PspA also inhibited C3 deposition when the classical pathway was initiated by antibodies to capsular polysaccharide . Antibodies to PspA could overcome the anticomplementary effect of PspA, allowing for increased complement activation and C3 deposition onto PspA+ bacteria.

Infect Immun, 2004 Jan, 72(1), 13 - 21
Further characterization of immunomodulation by a monoclonal antibody against Streptococcus mutans antigen P1; Rhodin NR et al.; We demonstrated previously that mucosal immunization of mice with Streptococcus mutans coated with the monoclonal antibody (MAb) 6-11A directed against the major surface adhesin protein P1 results in changes in the amount, isotype distribution, and specificity of serum antibodies compared with animals immunized with bacteria only . We now show that the specificity of the mucosal secretory IgA response was also influenced by this MAb . Changes in antibody specificity were associated with changes in biological activity . Serum samples which differed in antibody reactivity with P1 polypeptides generated by partial digestion with N-chlorosuccinimide but not in isotype distribution or overall reactivity with S . mutans or intact P1 demonstrated a statistically significant difference in the ability to inhibit bacterial adherence to salivary-agglutinin-coated hydroxyapatite beads . Serum IgG antibodies against P1 from mice immunized with either S . mutans alone or S . mutans coated with 6-11A were shown to recognize antigenic determinants dependent on the presence of the central proline-rich repeat domain, a segment necessary for the structural integrity of the molecule . However, no statistically significant differences were observed in antibody reactivity with a panel of six partial P1 polypeptides encoded by overlapping spaP subclones, suggesting that the targets of biologically relevant antibodies involve complex epitopes not reconstituted by the recombinant products tested . Lastly, we show that binding of MAb 6-11A to P1 on the surface of S . mutans alters P1's susceptibility to proteolytic digestion . Hence, changes in antigen processing and presentation may contribute to the immunomodulatory effects of this MAb.

J Am Geriatr Soc, 2003 Nov, 51(11), 1526 - 32
Ertapenem therapy for community-acquired pneumonia in the elderly; Woods GL et al.; OBJECTIVES: To compare the efficacy and safety of ertapenem, 1 g once a day, with ceftriaxone, 1 g once a day, for treatment of the subgroup of patients aged 65 and older with community-acquired pneumonia (CAP) requiring parenteral therapy . DESIGN: Combined data from patients aged 65 and older in two randomized, double-blind clinical trials . SETTING: Eighty international centers . PARTICIPANTS: Eight hundred fifty-seven treated patients, of whom 351 were aged 65 and older . INTERVENTIONS: Intravenous or intramuscular ertapenem or ceftriaxone with the option to switch to oral amoxicillin-clavulanate after at least 3 days of parenteral therapy . MEASUREMENTS: Clinical efficacy was assessed at completion of parenteral therapy and 7 to 14 days after all therapy had been completed (test of cure (TOC) assessment) . Bacterial eradication was assessed at the TOC visit . Safety was assessed daily during study therapy and for 14 days thereafter . RESULTS: One hundred forty-eight clinically evaluable patients aged 65 and older were treated with ertapenem and 125 with ceftriaxone . Pathogens were identified in 157 (57.5%) patients (the most common being Streptococcus pneumoniae), most of which were penicillin-susceptible . Clinical cure rates were 95.9% for patients in the ertapenem group and 92.7% for patients in the ceftriaxone group at completion of parenteral therapy and 93.9% and 90.4%, respectively, at the TOC assessment . Overall bacterial eradication rates were 92.8% (77 of 83) for patients treated with ertapenem and 93.2% (69 of 74) for those treated with ceftriaxone . The most common drug-related adverse experiences in both treatment groups were diarrhea and mild to moderate elevation of serum aminotransferase levels . CONCLUSION: Ertapenem 1 g once a day was highly effective for treatment of elderly patients with CAP requiring parenteral therapy and was as effective as ceftriaxone . Ertapenem was generally well tolerated, with an overall safety profile similar to ceftriaxone.

J Am Geriatr Soc, 2004 Jan, 52(1), 46 - 50
Healthy elderly people lack neutrophil-mediated functional activity to type V group B Streptococcus; Amaya RA et al.; OBJECTIVES: To determine the function of capsular polysaccharide (CPS)-specific immunoglobulin-G (IgG) and neutrophils from older adults in increasing ingestion and killing of type V group B Streptococcus (GBS) . DESIGN: Cross-sectional study . SETTING: Outpatient clinic at Baylor College of Medicine . PARTICIPANTS: The subjects were 40 healthy, community-dwelling adults aged 65 and older from Houston, Texas . MEASUREMENTS: The serum level of type V GBS CPS-specific IgG was measured using an enzyme-linked immunosorbent assay . Functional activity was evaluated using an opsonophagocytosis assay . RESULTS: Sera from four subjects promoted efficient neutrophil-mediated phagocytosis and killing of type V GBS (mean log10 reduction+/-standard deviation in colony-forming units (cfu)=1.51+/-0.39) . Each had serum CPS-specific IgG concentrations exceeding 1 microg/mL . Sera from 36 subjects did not promote neutrophil-mediated functional activity (mean log10 reduction in cfu=-0.09+/-0.06; P=.025) . Only one of these 36 had a CPS-specific IgG concentration exceeding 1 microg/mL . When pooled sera from young adults given type V GBS conjugate vaccine was added at CPS-specific IgG concentrations of 4 microg/mL or 0.4 microg/mL, sera from all subjects promoted neutrophil-mediated killing of type V GBS . No impairment was evident in the neutrophil function of elderly subjects when it was compared with that of young adults . CONCLUSION: CPS-specific IgG and neutrophils from healthy older adults function to ingest and kill type V GBS, but these antibodies are not present in sufficient amounts in most individuals . Further studies should determine whether a type V GBS vaccine induces functionally active antibodies in older people.

Eur J Biochem, 2004 Jan, 271(1), 14 - 22
Active site residues and mechanism of UDP-glucose dehydrogenase; Ge X et al.; UDP-glucose dehydrogenase catalyzes the NAD+-dependent twofold oxidation of UDP-glucose to give UDP-glucuronic acid . A sequestered aldehyde intermediate is produced in the first oxidation step and a covalently bound thioester is produced in the second oxidation step . This work demonstrates that the Streptococcus pyogenes enzyme incorporates a single solvent-derived oxygen atom during catalysis and probably does not generate an imine intermediate . The reaction of UDP-{6",6"-di-2H}-d-glucose is not accompanied by a primary kinetic isotope effect, indicating that hydride transfer is not rate determining in this reaction . Studies with a mutant of the key active site nucleophile, Cys260Ala, show that it is capable of both reducing the aldehyde intermediate, and oxidizing the hydrated form of the aldehyde intermediate but is incapable of oxidizing UDP-glucose to UDP-glucuronic acid . In the latter case, a ternary Cys260Ala/aldehyde intermediate/NADH complex is presumably formed, but it does not proceed to product as both release and hydration of the bound aldehyde occur slowly . A washout experiment demonstrates that the NADH in this ternary complex is not exchangeable with external NADH, indicating that dissociation only occurs after the addition of a nucleophile to the aldehyde carbonyl . Studies on Thr118Ala show that the value of kcat is reduced 160-fold by this mutation, and that the reaction of UDP-D-{6",6"-di-2H}-glucose is now accompanied by a primary kinetic isotope effect . This indicates that the barriers for the hydride transfer steps have been selectively increased and supports a mechanism in which an ordered water molecule (H-bonded to Thr118) serves as the catalytic base in these steps.

J Pediatr (Rio J), 1999 Jul, 75(Suppl 1), S103 - 14
{Streptococcal infections}; Santos VP; OBJECTIVES: To review the main diseases caused by the different strains of streptococcus, and to discuss the practical aspects of clinical diagnosis and the range of therapeutic treatments . METHODOLOGY: A review of the literature and a selection of the most meaningful themes for the pediatricians . RESULTS: The majority of the streptococcus described here are susceptible and have sensibility to penicillin . Different kinds of treatments aim at a better support to the offered scheme, including smaller amounts of doses per day, short-lived treatment and lowcost . CONCLUSION: Streptococcus infections are very frequently seen in the pediatrician's office . It is important to make an accurate diagnosis, and if possible a more simplified therapeutic treatment.

J Pediatr (Rio J), 1999 Jul, 75(Suppl 1), S74 - 90
{Penicillin-resistant pneumococcus: practical implications}; Mantese OC; OBJECTIVE: To review epidemiological, laboratorial and clinical aspects of the penicillin-resistant pneumococcus, and to consider the impact of the rapidly increasing drug-resistance on the management of the main infections caused by these pathogens . METHODS: Bibliographic review covering the last ten years, using both Medline system and direct research . RESULTS AND CONCLUSIONS: The Streptococcus pneumoniae is an important agent in acute otitis media, pneumonia and meningitis in children . The clinical evaluation of the pneumococcal infections depends on the age and health condition of the patient, site and severity of the infection and the adequacy of the treatment . Penicillin was uniformly effective against pneumococcus until three decades ago, when the first reports of clinical resistance were published . Since then, there has been a rapidly increasing in the level and rates of resistance to penicillin, which parallels to others beta lactams and antimicrobials . This paper includes a suscint review of the genetic and phenotypic mechanisms of the resistance to penicillin, the available bacteriologic tests for determination of in vitro susceptibility to antimicrobials and finally the importance of the pneumococcus in acute otitis media, pneumonia/bacteremia and meningitis . There has been new empirical therapeutic regimens recommended for meningitis, according to the community rates and levels of resistance to beta lactams and to the presence of certain underlying conditions among the patients, such as immunedepressant diseases and frequent antibiotic consumption . The rapidly increasing prevalence of drug-resistant strains emphasizes the importance of judicious antibiotic use and the development of a conjugate vaccine, effective to prevent invasive infections and to reduce the rate of carriage of the pneumococcus, even among infants.

J Pediatr (Rio J), 2003 Nov-Dec, 79(6), 537 - 42
{Prevalence of serotypes and antimicrobial resistance of invasive strains of Streptococcus pneumoniae}; Mantese OC et al.; OBJECTIVE: To determine the prevalence of serotypes and antimicrobial susceptibility of invasive strains of Streptococcus pneumoniae and to discuss the implications of these findings for vaccine formulation . METHODS: Strains of Streptococcus pneumoniae obtained from normally sterile fluids from patients admitted with invasive diseases were isolated and identified at the Hospital de Clinicas, Universidade Federal de Uberlandia, state of Minas Gerais, and forwarded to Instituto Adolfo Lutz, state of Sao Paulo, for further identification, serotyping and determination of antimicrobial susceptibility . RESULTS: From April 1999 to March 2003, 148 invasive strains of Streptococcus pneumoniae were obtained . The age of patients ranged from 1 day to 88.83 years (mean: 21.33+/-25.82 years; median: 4.42 years) . Eighty-four (56.7%) patients were male . The most common diagnoses were pneumonia (91 cases; 61.4%), meningitis (32 cases; 21.6%) and occult bacteremia (15 cases; 10.1%) . Strains were isolated mostly from blood (76 occasions; 51.3%), pleural fluid (39 occasions; 26.3%) and cerebrospinal fluid (30 occasions; 20.2%) . There were 23 different serotypes, and the most common were 14, 3, 1, 5, 6A, 6B and 18C . Among 30 (20.2%) oxacillin-resistant strains, 23 (15.5%) were confirmed as resistant to penicillin (12.8% intermediate resistance and 2.7% full resistance) . Oxacillin-resistant strains were restricted to serotypes 14, 23F, 19A and 6B . Resistance to penicillin varied with age, being more common in children under two years of age (p = 0.0008) . We observed decreased sensibility to sulfamethoxazole-trimethoprim (92 isolates; 63.4%), to erythromycin (12 isolates; 8.3%), to clindamycin (12 isolates; 8.7%), to ofloxacin (one strain; 0.8%) and to cefotaxime (three strains; 2%; also resistant to penicillin) . All isolates were susceptible to chloramphenicol, rifampin and vancomycin . CONCLUSIONS: The decreased susceptibility to penicillin, detected in 15.5% of the strains was predominant in children under two years of age . There were 23 different Streptococcus pneumoniae serotypes . The 23-valent polysaccharide vaccine covers 82.6% of the serotypes and 90.2% of the invasive strains isolated in this population . In addition, 46.7% of the serotypes and 63.6% of the strains isolated from children until five years of age are covered in the currently available 7-valent conjugated vaccine (PN CRM7).

J Vet Sci, 2003 Dec, 4(3), 213 - 24
Identification and epidemiological characterization of Streptococcus uberis isolated from bovine mastitis using conventional and molecular methods; Khan IU et al.; In the present study 130 S . uberis strains and one S . parauberis strain isolated from bovine milk samples of 58 different farms of various locations in Hesse, Germany, as well as two reference strains of each species were comparatively investigated for cultural, biochemical, serological and molecular properties . All S . uberis strains produced the enzyme beta-D-glucuronidase, while the S . parauberis strains were negative . The S . uberis and S . parauberis 16S rRNA genes were amplified by polymerase chain reaction and subsequently digested with the restriction enzymes RsaI and AvaII yielding species-specific restriction patterns . Both species were additionally identified by amplifying species-specific parts of the genes encoding the 16S rRNA, the 23S rRNA and the 16S-23S rDNA intergenic spacer region, respectively . The CAMP factor gene cfu, a potential virulence factor of S . uberis, was amplified, corresponding to a phenotypically positive CAMP-reaction, using cfu-specific oligonucleotide primers . In addition the streptokinase/plasminogen activator encoding genes skc/pauA, a second potential virulence factor, could be amplified for 126 of the 130 S . uberis but not for S . parauberis . A DNA fingerprinting of S . uberis strains, performed by macrorestriction analysis of their chromosomal DNA by pulsed-field gel electrophoresis, revealed that most of the isolates were not related to each other . However, identical DNA patterns were noted for some of the isolates within different quarters of an individual cow and also for different cows within the same farm . The generally unrelated DNA patterns indicated that S . uberis is a pathogen with multiple environmental habitats and that infections are caused by a great variety of strains.

Caries Res, 2004 Jan-Feb, 38(1), 9 - 14
The effects of egg-derived antibodies to glucosyltransferases on dental caries in rats; Kruger C et al.; The role of Streptococcus mutans in the development of dental caries is well recognized . Important virulence factors include the glucosyltransferases (gtf), essential for production of glucans . We evaluated the anticariogenic effects of orally administered chicken anti-cell-associated (CA) Gtf antibodies in desalivated rats . The animals were infected with S . mutans MT8148R and treated with chicken anti-CA-Gtf egg yolk antibodies (IgY) or nonimmune egg yolk powder . Smooth surface lesions were significantly lower in the anti-CA-Gtf-treated group in comparison to the control groups . Sulcal surface caries was also decreased and of less severity . Our study suggests that chicken anti-CA-Gtf antibodies may have promise as a prophylaxis for high caries risk patients .

Lancet, 2003 Dec 13, 362(9400), 1991 - 2001
Community-acquired pneumonia; File TM; This seminar reviews important features and management issues of community-acquired pneumonia (CAP) that are especially relevant to immunocompetent adults in light of new information about cause, clinical course, diagnostic testing, treatment, and prevention . Streptococcus pneumoniae remains the most important pathogen; however, emerging resistance of this organism to antimicrobial agents has affected empirical treatment of CAP . Atypical pathogens have been quite commonly identified in several prospective studies . The clinical significance of these pathogens (with the exception of Legionella spp) is not clear, partly because of the lack of rapid, standardised tests . Diagnostic evaluation of CAP is important for appropriate assessment of severity of illness and for establishment of the causative agent in the disease . Until better rapid diagnostic methods are developed, most patients will be treated empirically . Antimicrobials continue to be the mainstay of treatment, and decisions about specific agents are guided by several considerations that include spectrum of activity, and pharmacokinetic and pharmacodynamic principles . Several factors have been shown to be associated with a beneficial clinical outcome in patients with CAP . These factors include administration of antimicrobials in a timely manner, choice of antibiotic therapy, and the use of a critical pneumonia pathway . The appropriate use of vaccines against pneumococcal disease and influenza should be encouraged . Several guidelines for management of CAP have recently been published, the recommendations of which are reviewed.

FEMS Microbiol Lett, 2003 Dec 12, 229(2), 179 - 82
Characterization of a suppressor mutation complementing an acid-sensitive mutation in Streptococcus mutans; Lis M et al.; We isolated a spontaneous suppressor mutant complementing the acid-sensitive phenotype of Streptococcus mutans strain Tn-1, a mutant previously generated in this laboratory, defective in the activity of the dgk-encoded putative undecaprenol kinase . A relatively simple genetic method was developed to identify the suppressor mutation, based on selection for transformants containing two closely linked markers: a selectable allele of the unknown suppressor gene and an antibiotic resistance gene introduced on a suicide plasmid at random sites into the chromosome via homologous recombination . While we have not actually identified the original suppressor mutation, another mutated gene restoring acid resistance has been isolated, which suggests a possible mechanism of suppression.

Zhonghua Kou Qiang Yi Xue Za Zhi, 2003 Sep, 38(5), 342 - 4
{Adhesion of oral microorganisms on dental porcelain polished and glazed}; Wang YN et al.; OBJECTIVE: This study compared the roughness of porcelain polished or glazed surfaces and the adhesion of oral streptococcus mutans to them in vitro . METHODS: 30 porcelain samples were made . Porcelain samples in group A were polished with diamond paste . Porcelain samples were glazed in group B and were polished with Al2O3 (240#) bur in group C . Their roughness values were measured by profilometer . Standardized cell suspensions were incubated with test samples for one hour at 37 degrees C, then retained cells were counted by image analysis (percentage area of a microscopic field covered by cells) . RESULTS: Roughness values of group A, B, C were respectively (0.1987 +/- 0.057) microm, (0.1990 +/- 0.091) microm, (0.4260 +/- 0.174) microm . There was no significantly difference between group A and group B . The roughness samples in group C were significantly rougher than that in the other groups . The amount of retained cells in group A, group B, group C was respectively (15.92 +/- 4.37)%, (16.39 +/- 6.31)% and (41.48 +/- 12.1)% . There was no significant difference between the cell adhesion on porcelain surface glazed and polished, but more bacteria adhered on the porcelain surface in group C . CONCLUSIONS: Porcelain surface polished treatment was clinically acceptable compared with its glazed . They all exhibited the least amount of bacteria adhesion . The more porcelain surface was rough, the more bacteria adhered on it.

Clin Infect Dis, 2004 Jan 1, 38(1), 99 - 103 Epub 2003 Dec 08.
The clinical significance of macrolide-resistant Streptococcus pneumoniae: it's all relative; Nuermberger E et al.; Macrolides are currently recommended as first-line agents for the empirical treatment of community-acquired pneumonia . Heavy use of these agents for a variety of indications has resulted in an increasing incidence of macrolide resistance among pneumococcal isolates . Although several case reports and small case series have suggested that in vitro macrolide resistance is associated with treatment failure in cases of pneumococcal pneumonia, other observational data suggest that drug susceptibility testing may not correlate with treatment failure . In this article, we review current information on the mechanisms of macrolide resistance and the pharmacodynamics of macrolide therapy, together with efficacy data from animal models and clinical observations, to begin to gauge the clinical significance of macrolide resistance in Streptococcus pneumoniae . Areas for further investigation are highlighted.

J Bacteriol, 2004 Jan, 186(1), 110 - 21
Natural selection and evolution of streptococcal virulence genes involved in tissue-specific adaptations; Kalia A et al.; The molecular mechanisms underlying niche adaptation in bacteria are not fully understood . Primary infection by the pathogen group A streptococcus (GAS) takes place at either the throat or the skin of its human host, and GAS strains differ in tissue site preference . Many skin-tropic strains bind host plasminogen via the plasminogen-binding group A streptococcal M protein (PAM) present on the cell surface; inactivation of genes encoding either PAM or streptokinase (a plasminogen activator) leads to loss of virulence at the skin . Unlike PAM, which is present in only a subset of GAS strains, the gene encoding streptokinase (ska) is present in all GAS isolates . In this study, the evolution of the virulence genes known to be involved in skin infection was examined . Most genetic diversity within ska genes was localized to a region encoding the plasminogen-docking domain (beta-domain) . The gene encoding PAM displayed strong linkage disequilibrium (P << 0.01) with a distinct phylogenetic cluster of the ska beta-domain-encoding region . Yet, ska alleles of distant taxa showed a history of intragenic recombination, and high intrinsic levels of recombination were found among GAS strains having different tissue tropisms . The data suggest that tissue-specific adaptations arise from epistatic coselection of bacterial virulence genes . Additional analysis of ska genes showed that approximately 4% of the codons underwent strong diversifying selection . Horizontal acquisition of one ska lineage from a commensal Streptococcus donor species was also evident . Together, the data suggest that new phenotypes can be acquired through interspecies recombination between orthologous genes, while constrained functions can be preserved; in this way, orthologous genes may provide a rich and ready source for new phenotypes and thereby play a facilitating role in the emergence of new niche adaptations in bacteria.

Oral Microbiol Immunol, 2004 Feb, 19(1), 57 - 60
Inactivation of srtA gene of Streptococcus mutans inhibits dextran-dependent aggregation by glucan-binding protein C; Igarashi T et al.; A sortase-deficient mutant of Streptococcus mutans was prepared by insertional inactivation of a sortase gene (srtA) . The srtA mutant was defective in cell wall-anchoring of two surface proteins 200 and 75 kDa in size . A previous study has shown that the 200 kDa protein is a surface protein antigen PAc and that the sortase catalyzes cell wall-anchoring of PAc in S . mutans . In this study another surface protein 75 kDa in size was examined by immunologic and physiologic methods . Western blot analysis with a specific antiserum showed that the 75 kDa protein was a surface protein, glucan-binding protein C . The protein was overexpressed under a stress condition including a sublethal concentration of tetracycline . The srtA mutant cells also lost the ability of dextran-dependent aggregation . These results suggest that the S . mutans sortase mediates cell wall-anchoring of the glucan-binding protein C and dextran-dependent aggregation of this organism.

Cell Microbiol, 2004 Jan, 6(1), 79 - 88
Enolase from Streptococcus sobrinus is an immunosuppressive protein; Veiga-Malta I et al.; A strategy of Streptococcus sobrinus, a major agent of dental caries, to survive and colonize the host consists of the production of a protein that suppresses the specific antibody responses . We have cloned the gene coding for a protein with immunosuppressive activity . It contains an open reading frame of 1302 base pairs encoding a polypeptide with 434 amino acid residues and a molecular mass of 46910 Da . The gene product is homologous to enolases from several organisms . The polypeptide was expressed in Escherichia coli as a hexahistidine-tagged protein and purified in a fluoride-sensitive enzymatically active form . Pretreatment of mice with the S . sobrinus recombinant enolase suppresses a primary immune response against T-cell dependent antigens . This immunosuppressive effect is specific to the antigen used in the immunization, as it is not observed when the immune response against other antigens is analysed . Furthermore, the S . sobrinus recombinant enolase stimulates an early production of interleukin-10, an anti-inflammatory cytokine, and not the pro-inflammatory cytokine IFN-gamma . These observations indicate that enolase acts in the suppression of the specific host immune response against S . sobrinus infection.

Int J Tuberc Lung Dis, 2003 Dec, 7(12), 1207 - 8
Dual infection with Streptococcus pneumoniae and Mycobacterium tuberculosis in HIV-seropositive patients with community acquired pneumonia; Schleicher GK et al.; Pulmonary infections with more than one organism are common in human immunodeficiency virus (HIV) seropositive patients . We describe nine cases of dual infection with Streptococcus pneumoniae and Mycobacterium tuberculosis in HIV-seropositive patients presenting with community acquired pneumonia (CAP) . It is important to exclude pulmonary tuberculosis in HIV-seropositive patients with CAP who fail to respond appropriately to initial antibiotic therapy, even if another etiological pathogen has been found.

Acta Medica (Hradec Kralove), 2003, 46(3), 125 - 7
Infective endocarditis caused by group G streptococcus with multiple cerebral emboli; Erdem I et al.; The group G streptococcal endocarditis is a rare form of infective endocarditis . In this form of infective endocarditis, serious neurological complications most commonly develop . We reported this case because of its being an unusual form of infective endocarditis that was caused by Group G Streptococcus . We also reviewed the literature . The patient was admitted to infectious disease service with a presumptive diagnosis of central nervous system infection . Blood cultures were positive for group G streptococcus . There was a mass on the posterior surface of the mitral valve which was 2 x 2.5 cm in length on the echocardiography . In the cranial computerized tomography of our patient, slightly increased contrast media uptake was observed in the both parietal lobes, in the both frontal lobes, and in the anterior areas of right occipital lobe . Therefore, this case was assumed as infective endocarditis caused by group G streptococcus with multiple cerebral emboli . Ceftriaxone was given for 4 weeks and gentamicin was given for 2 weeks, and progressive improvement of the patient's condition was seen.

Arch Microbiol, 2004 Feb, 181(2), 122 - 8 Epub 2003 Dec 16.
Molecular characteristics and transcription of the gene encoding a multifunctional alcohol dehydrogenase in relation to the deactivation of pyruvate formate-lyase in the ruminal bacterium Streptococcus bovis; Asanuma N et al.; To clarify the deactivation mechanism of pyruvate formate-lyase (PFL) and its role in the regulation of fermentation in Streptococcus bovis, the molecular properties and genetic expression of multifunctional alcohol dehydrogenase (ADHE) were investigated . S . bovis was found to have ADHE, which was deduced to consist of 872 amino acids with a molecular mass of 97.4 kDa . The ADHE was shown to harbor three enzyme activities: (1) alcohol dehydrogenase, (2) coenzyme-A-linked acetaldehyde dehydrogenase that catalyzes the conversion of acetyl-CoA to ethanol, and (3) PFL deactivase . Similar to Escherichia coli ADHE, S . bovis ADHE required Fe2+ for its activity . The gene encoding ADHE ( adhE) was shown to be monocistronic . The level of adhE mRNA changed in parallel with the mRNA levels of the genes encoding PFL (pfl) and PFL-activating enzyme (act) as the growth conditions changed, although these genes are independently transcribed . Synthesis of ADHE, PFL-activating enzyme, and PFL appears to be regulated concomitantly . Overexpression of ADHE did not cause a change in the formate-to-lactate ratio . It is conceivable that ADHE is not significantly involved in the reversible inactivation of active PFL under anoxic conditions . Partition of the flow from pyruvate appears to be mainly regulated by the activities of lactate dehydrogenase and PFL.

Arch Microbiol, 2004 Feb, 181(2), 106 - 11 Epub 2003 Dec 16.
Purification of saliva agglutinin of Streptococcus intermedius and its association with bacterial aggregation and adherence; Yamaguchi T; Streptococcus intermedius strain 1208-1 cells were aggregated in the presence of saliva . The saliva agglutinin was purified by centrifugation, filtration, and gel filtration . SDS-PAGE analyses indicated that the purified agglutinin consisted of two high-molecular-mass proteins . Aggregation was dependent on calcium over pH 5.5, with 1 mM being the most effective concentration . Boiling inactivated purified agglutinin . S . intermedius strain 3 and Streptococcus mutans strain 1 were aggregated in the purified agglutinin . After adsorption with strain 1208-1 cells, the saliva sample did not exhibit any aggregation activity, and the agglutinin bands were no longer visible by SDS-PAGE . Adherence analyses demonstrated that the purified agglutinin immobilized on the surfaces of polystyrene wells, actinomyces cells, and apatite beads accounted for the binding of streptococcus cells . Agglutinin also effectively inhibited adherence to apatite beads coated with native saliva.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2003 Dec, 96(6), 751 - 6
Reduction of viable bacteria in dentinal tubules treated with clindamycin or tetracycline; Lin S et al.; AIM: We sought to evaluate and compare the antibacterial effect of clindamycin and tetracycline in bovine dentinal tubules . METHODS: Dentinal tubules of 32 cylindrical bovine root specimens were infected with Streptococcus sanguis N1 . Clindamycin 2% or tetracycline 2% (Ledermix) was placed in the root canal for 1 week . Powder dentin samples obtained from within the canal lumina by using International Standards Organization No . 25 to No . 31 burs were examined for the presence of vital bacteria after the brain-heart infusion plates were inoculated and the colony-forming units were counted . The potent effect of the medicaments was also evaluated through the use of the agar diffusion test . RESULTS: Heavy bacterial infection was observed in the control bovine root specimens at the layer close to the lumen . This decreased rapidly from layer to layer up to the deepest layer tested (300-400 microm), which contained several hundred colony-forming units . Clindamycin significantly reduced the amount of viable bacteria in each dentin layer compared with the positive control and tetracycline (P <.01) . The agar diffusion test, wherein dilutions in increments of 1/3 and 1/9 were used, revealed that both medicaments had antibacterial activity, but clindamycin was significantly better . In the 1/27 dilution, clindamycin had a minor effect and tetracycline had no effect at all . CONCLUSION: Under the experimental conditions used in this study, the commercial preparations of clindamycin were more effective than those of tetracycline (Ledermix) in the agar diffusion test and clindamycin penetrated into dentinal tubules up to 400 microm . Thus, it has the potential to serve as an effective intracanal medicament in persistent infections when other medicaments fail.

J Orthop Surg (Hong Kong), 2003 Dec, 11(2), 221 - 3
Group G streptococcus--a rare cause of osteomyelitis simulating bone tumour: a case report; Tong SH et al.; We report a case of osteomyelitis of the proximal femur caused by Lancefield group G streptococcus in a 71-year-old otherwise healthy man . The organism has rarely been identified as the cause of osteomyelitis . The subacute nature of the symptoms and the radiological appearance of the femur in this patient mimic bone tumour . The patient was successfully treated with conservative methods, including a prolonged period of oral antibiotics . We stress the importance of histological and bacteriological evidence in avoiding misdiagnosing patients with equivocal clinical and radiological presentation.

J Infect Dis, 2003 Dec 15, 188(12), 1928 - 39 Epub 2003 Dec 03.
Effects of rhinovirus infection on the adherence of Streptococcus pneumoniae to cultured human airway epithelial cells; Ishizuka S et al.; To examine the effects of rhinovirus (RV) infection on the adherence of Streptococcus pneumoniae to human tracheal epithelial cells, cells were infected with RV-14, and S . pneumoniae were added to the culture medium . The number of S . pneumoniae adhering to epithelial cells increased after RV infection . Y-24180, a specific inhibitor of the platelet-activating factor receptor (PAF-R); PAF; and the pyrrolidine derivative of dithiocarbamate, an inhibitor of transcription factor nuclear factor-kappaB (NF-kappaB), decreased the number of S . pneumoniae adhering to cells after RV-14 infection . RV-14 infection increased PAF-R expression and the activation of NF-kappaB and promoter-specific transcription factor 1 . These findings suggest that RV-14 infection stimulates S . pneumoniae adhesion to airway epithelial cells via increases in PAF-Rs that are partly mediated through activation of transcription factors . Increased adherence of S . pneumoniae may be one of the reasons that pneumonia develops after RV infection.

J Infect Dis, 2003 Dec 15, 188(12), 1898 - 908 Epub 2003 Dec 09.
Structure and distribution of an unusual chimeric genetic element encoding macrolide resistance in phylogenetically diverse clones of group A Streptococcus; Banks DJ et al.; The resistance of group A Streptococcus (GAS) to macrolide antibiotics is now a worldwide problem . Preliminary sequencing of the genome of an erythromycin-resistant serotype M6 clone that was responsible for a pharyngitis outbreak in Pittsburgh, Pennsylvania, was conducted to determine the structure of the genetic element containing the mefA gene, which encodes a macrolide efflux protein . The mefA gene is associated with a 58.8-kb chimeric genetic element composed of a transposon inserted into a prophage . This element also encodes a putative extracellular protein with a cell-wall anchoring motif (LPKTG) located at the carboxyterminus . The mefA element was present in phylogenetically diverse GAS strains isolated throughout the United States . Culture supernatants, prepared after mitomycin C treatment, of a strain representing the outbreak clone contained mefA element DNA in a DNAse-resistant form . Together, these data provide new information about the molecular genetic basis of macrolide resistance and dissemination in GAS strains.

J Biol Chem, 2004 Feb 27, 279(9), 8252 - 61 Epub 2003 Dec 14.
The YefM antitoxin defines a family of natively unfolded proteins: implications as a novel antibacterial target; Cherny I et al.; Although natively unfolded proteins are being observed increasingly, their physiological role is not well understood . Here, we demonstrate that the Escherichia coli YefM protein is a natively unfolded antitoxin, lacking secondary structure even at low temperature or in the presence of a stabilizing agent . This conformation of the protein is suggested to have a key role in its physiological regulatory activity . Because of the unfolded state of the protein, a linear determinant rather than a conformational one is presumably being recognized by its toxin partner, YoeB . A peptide array technology allowed the identification and validation of such a determinant . This recognition element may provide a novel antibacterial target . Indeed, a pair-constrained bioinformatic analysis facilitated the definite determination of novel YefM-YoeB toxin-antitoxin systems in a large number of bacteria including major pathogens such as Staphylococcus aureus, Streptococcus pneumoniae, and Mycobacterium tuberculosis . Taken together, the YefM protein defines a new family of natively unfolded proteins . The existence of a large and conserved group of proteins with a clear physiologically relevant unfolded state serves as a paradigm to understand the structural basis of this state.

BMC Infect Dis . 2003 Dec 12;3(1):29.
HIV infected adults do not have an excess of colonising bacteria in their large airways; Gordon SB et al.; BACKGROUND: HIV infected adults have increased susceptibility to bacterial pneumonia but the underlying immune defect is poorly understood . We tested the hypothesis that HIV infection might be associated with increased bacterial colonisation of distal airways by nasal flora, which would then predispose patients to bacterial pneumonia . METHODS: Healthy volunteer adults with normal chest radiographs were recruited . Bronchoscopy was carried out and uncontaminated mucosal samples were collected from proximal and distal sites in the large airways using a protected specimen brush . Samples were cultured to detect typical respiratory tract colonising organisms, and the proportion of samples found to contain colonising bacteria compared between HIV infected and uninfected subjects using non-parametric tests . RESULTS: Forty-nine subjects were studied of whom 27 were HIV infected . Colonising bacteria were identified in the nasopharynx of all subjects including Streptococcus pneumoniae in 6/49 subjects (5 HIV uninfected) . Colonising bacteria were found in the distal airway of 6 subjects (3/27 HIV infected vs 3/22 HIV uninfected; chi2 = 0.07, p = 0.8) . Streptococcus pneumoniae was identified in the trachea of all subjects with nasal colonisation but in the distal airway of only 1 subject . CONCLUSIONS: There was no evidence to support a hypothesis of increased airway bacterial colonisation in healthy HIV infected subjects.

J Dairy Sci, 2003 Nov, 86(11), 3783 - 6
Efficacy of two iodine teat dips during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae; Foret CJ et al.; An experimental challenge trial was performed against Staphylococcus aureus and Streptococcus agalactiae following the procedures recommended by the National Mastitis Council . The efficacy of two teat dips, product 1 (Bovadine with I-Tech II) and product 2 (Bovadine with I-Tech, used as a positive control), was determined . Both teat dips contain 1% iodine and 10% glycerin . Product 1 established an 89.7% reduction in infections against Staph . aureus and 73.1% reduction in infections against Strep . agalactiae . Product 2 demonstrated an 86.2% reduction in infections against Staph . aureus and 78.4% reduction in infections against Strep . agalactiae . Teat skin and teat ends were evaluated before and after the trial . No significant change in teat condition was observed for either product.

J Dairy Sci, 2003 Nov, 86(11), 3466 - 72
Results of milk samples submitted for microbiological examination in Wisconsin from 1994 to 2001; Makovec JA et al.; The objective of this study was to examine the characteristics of milk samples submitted for microbiological examination at the Wisconsin Veterinary Diagnostic Laboratory between 1994 and 2001 . Results (n = 83,650) of microbiological testing of milk samples (n = 77,172) submitted to the Wisconsin Veterinary Diagnostic Laboratory from January 1994 until June 2001 were analyzed . Submissions included milk samples obtained from cases of clinical and subclinical mastitis as well as samples obtained for mastitis surveillance programs . Results were recorded as no growth, contaminated, or identified as specific bacterial pathogens . Statistical analysis was performed to determine trends in the isolation of mastitis pathogens . The proportion of samples identified as contaminated decreased from 20.6 (1997) to 9.5% (2001) . The proportion of samples coded as no growth increased from 22.6 (1994) to 49.7% (2001) . Isolation of Staphylococcus aureus decreased from 17.7% (1994) of isolates to 9.7% (2001), while isolation of Streptococcus agalactiae decreased from 8.1 (1994) to 3.0% (2001) . Coagulase-negative Staphylococcus spp . were isolated from 12.7 to 17.5%, environmental Streptococcus spp . were isolated from 11.6 to 20.1%, and Escherichia coli were isolated from 3.1 to 6.7% of all isolates . No growth and contaminated samples comprised almost 50% of total submissions, and it is important that producers have proper expectations when submitting milk samples . The proportion of isolates identified as Staph . aureus and Strep . agalactiae decreased, suggesting the proportion of contagious bacteria causing mastitis has decreased . Environmental and contagious pathogens demonstrated characteristic differences by season.

Arch Dis Child, 2003 Dec, 88(12), 1112 - 8
The effect of HIV infection on paediatric bacterial meningitis in Blantyre, Malawi; Molyneux EM et al.; AIM: To compare presentation, progress, and outcome of acute bacterial meningitis in HIV seropositive and seronegative children . METHODS: A double blind randomised placebo controlled study of the use of dexamethasone as adjuvant therapy in acute bacterial meningitis, in children aged 2 months to 13 years, was carried out from July 1997 to March 2001 . A total of 598 children were enrolled, of whom 459 were tested for HIV serostatus . RESULTS: Of the 459 children, 34% were HIV seropositive . Their presentation was similar to HIV seronegative children but more were shocked on arrival at hospital (33/157 v 12/302), and more had a focus of infection (85/157 v 57/302) . HIV positive children had a higher incidence of Streptococcus pneumoniae infections (52% v 32%) . Sixty four cases relapsed; 67% were in HIV positive patients . The mortality in HIV positive children was 65% compared with 36% in HIV negative children . The number of survivors in each group was similar . Hearing loss was more common in HIV negative than HIV positive children (66.3% v 47.2%) . Steroid therapy had no influence on meningitis in HIV positive children, but the mortality in HIV negative children was 61% in children given steroids, and 39% in those who did not receive steroids . CONCLUSION: HIV seropositive children who develop bacterial meningitis have a high mortality and are prone to recurrent disease . There is an urgent need to prevent both primary and recurrent infections.

Carbohydr Res, 2003 Nov 14, 338(23), 2629 - 51
Chemo-enzymatic synthesis of tetra-, penta-, and hexasaccharide fragments of the capsular polysaccharide of Streptococcus pneumoniae type 14; Joosten JA et al.; The chemo-enzymatic synthesis is described of beta-D-Glcp-(1-->6)-{beta-D-Galp-(1-->4)}-beta-D-GlcpNAc-(1-->3)-beta-D-Galp-(1-->O(CH(2))(6)NH(2) (1), beta-D-Glcp-(1-->6)-{beta-D-Galp-(1-->4)}-beta-D-GlcpNAc-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->O(CH(2))(6)NH(2) (2), beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->O(CH(2))(6)NH(2) (3), and beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->6)-{beta-D-Galp-(1-->4)}-beta-D-GlcpNAc-(1-->O(CH(2))(6)NH(2) (4), representing fragments of the repeating unit of the Streptococcus pneumoniae serotype 14 capsular polysaccharide . Linear intermediate oligosaccharides 5-8 were synthesized via chemical synthesis, followed by enzymatic galactosylation using bovine milk beta-1,4-galactosyltransferase as a catalyst . The title oligosaccharides form suitable compounds for conjugation with carrier proteins, to be tested as potential vaccines in animal models.

Carbohydr Res, 2003 Nov 14, 338(23), 2611 - 27
Chemo-enzymatic synthesis of a tetra- and octasaccharide fragment of the capsular polysaccharide of Streptococcus pneumoniae type 14; Joosten JA et al.; The chemo-enzymatic synthesis is described of tetrasaccharide beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->6)-{beta-D-Galp-(1-->4)}-beta-D-GlcpNAc-(1-->O(CH(2))(6)NH(2) (1) and octasaccharide beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->6)-{beta-D-Galp-(1-->4)}-beta-D-GlcpNAc-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->6)-{beta-D-Galp-(1-->4)}-beta-D-GlcpNAc-(1-->O(CH(2))(6)NH(2) (2), representing one and two tetrasaccharide repeating units of Streptococcus pneumoniae serotype 14 capsular polysaccharide . In a chemical approach, the intermediate linear trisaccharide 3 and hexasaccharide 4 were synthesized . Galactose residues were beta-(1-->4)-connected to the internal N-acetyl-beta-D-glucosamine residues by using bovine milk beta-1,4-galactosyltransferase . Both title oligosaccharides will be conjugated to carrier proteins to be tested as potential vaccines in animal models.

Carbohydr Res, 2003 Nov 14, 338(23), 2605 - 9
Synthesis of tetra- and pentasaccharides corresponding to the capsular polysaccharide of Streptococcus pneumoniae type 9A&L, 9N and 9A; Alpe M et al.; Two tetrasaccharides, alpha-D-GlcAp-(1-->3)-alpha-D-Galp-(1-->3)-beta-D-ManpNAc-(1-->4)-beta-D-Glcp and alpha-D-GlcAp-(1-->3)-alpha-D-Glcp-(1-->3)-beta-D-ManpNAc-(1-->4)-beta-D-Glcp (protected form), and a pentasaccharide, alpha-D-Glcp-(1-->4)-alpha-D-GlcAp-(1-->3)-alpha-D-Galp-(1-->3)-beta-D-ManpNAc-(1-->4)-beta-D-Glcp have been synthesised from 2-aminoethyl glycoside trisaccharide acceptors in a linear approach via consecutive alpha-glycosylations . Ethyl thioglycosides were used as glycosyl donors and DMTST in Et(2)O or NIS/TfOH in CH(2)Cl(2) were employed as promoters.

Ginekol Pol, 2003 Oct, 74(10), 1223 - 7
{Prevalence of group B streptococcal colonization in pregnant women and their newborns based on the results of examination of patients in the Obstetric and Gynecology Department of the National Research Institute of Mother and Child--a pilot study}; Kowalska B et al.; Group B streptococcus is a main cause of perinatal infections and neonatal sepsis . In Poland there is no epidemiological data of the prevalence group B streptococcal colonization in pregnant women and the risk for their newborns . OBJECTIVES: The aim of the study is to define the prevalence of streptococcal B colonization in pregnant women and their newborns . MATERIALS AND METHODS: The pregnant women and their newborns from Obstetric and Gynecology Department of National Research Institute of Mother and Child were included to our study during 2001 and 2002 years . Cervical, vaginal and perianal swab were obtained . Women with positive cultures were treated with antibiotic during labor . The external swabs of their neonate were obtained . RESULTS: 1678 pregnant women took part in our study . The GBS (streptococcus agalactiae) colonization was found in 331 women . The prevalence of pregnant women group B streptococcal colonization in the study is 19.7% . 70 of 203 neonates form mothers with positive results of our screening, had the GBS colonization confirmed . The prevalence of confirmed streptococcal colonization in neonates was 34.5% . One newborn developed early onset neonatal sepsis, during the study period . CONCLUSIONS: The prevalence of pregnant women group B streptococcal colonization about 20% . For the prevention of newborns intrapartum infections a major thing is the prevalence of the transmission risk to newborns from mothers with a GBS colonization and the appropriate intrapartum management.

Arch Inst Pasteur Tunis, 1999 Jan-Apr, 76(1-4), 13 - 8
{Efficacy of intra-lesional glucantime in the treatment of zoonotic cutaneous leishmaniasis in basic health care conditions}; Chahed MK et al.; A randomized placebo-controlled trial treating cutaneous lesions due to Leishmania major with intralesionnel glucantime, was conducted in El Guettar between december 1994 and June 1995, in order to assess efficacy of this therapy under field conditions . It included 109 patients: 52 were administrated glucantime and 57 received local treatment (eosin 5% and alcohol 95%) . Prognostic factors were similar in both groups . Results did not reveal a significant difference between glucantime and eosin regarding the rapidity of the healing of lesions . However, scars seem to be of better quality among the glucantime group . Bacterial super infection was noticed among 57.6% of humid lesions sampled among 33 patients . Isolated strains included group A streptococcus (22%), staphylococcus aureus (16.7%) or an association of both agents (61.1%) . Resistance profile indicated that streptococcus and staphylococcus respond well to macrolids compared to other antibiotic groups.

Ann Epidemiol, 2004 Jan, 14(1), 44 - 8
Etiology, antimicrobial susceptibility profiles, and mortality associated with bacterial meningitis among children in Egypt; Youssef FG et al.; PURPOSE: Surveillance for patients with meningitis is a high priority in order to determine the etiology of disease and design prevention strategies . This study presents data on the causes of bacterial meningitis among children <6 years of age treated in a network of hospitals throughout Egypt . METHODS: Training was provided to standardize the collection of clinical information and optimize recovery of bacterial pathogens . Bacterial isolates were tested for antimicrobial resistance patterns using Kirby Bauer disk diffusion, E-test and/or Beta-lactamase (BL) testing methods . RESULTS: Patients with culture-confirmed bacterial meningitis (228 children<6 years) were identified including 89 (39%) patients with H . influenzae (HI), 68 (30%) with Streptococcus pneumoniae (SP), 30 (13%) with N . meningitidis (NM), 18 (8%) with Mycobacterium tuberculosis (MTB) and 23 (10%) with other bacteria . The overall case fatality ratio was high (24%) and increased among children with TB meningitis (56%) . The susceptibility for HI to ampicillin (AMP), chloramphenicol (C) and ceftriaxone (CRO) was 21%, 13%, and 100% respectively . The susceptibility for SP to C and CRO was 79% and 100%, respectively . CONCLUSIONS: HI and SP are the leading causes of bacterial meningitis among children in Egypt . The majority of HI strains tested were resistant to AMP or C suggesting the need for routine use of CRO as first line therapy . Among older children TB emerges as a significant cause of bacterial meningitis in Egypt.

Otolaryngol Head Neck Surg, 2003 Dec, 129(6), 666 - 73
Hearing loss with stapedotomy in otitis media; Falcone MT et al.; OBJECTIVE: Stapes fixation is often found in patients with either active or inactive otitis media (OM) . Stapedotomy is generally not performed in ears with OM because of the potential risk of sensorineural hearing loss (SNHL) . Recent studies have demonstrated that the inner ear may be fenestrated in the presence of OM without significantly increased risk of SNHL . The goal of this study was to determine if stapedotomy in the presence of OM significantly increases the incidence of postoperative SNHL . METHODS: Streptococcus pneumoniae or Pseudomonas aeruginosa was injected bilaterally into the middle ears of guinea pigs (n = 18 and 16, respectively) . A control group (n = 14) was injected with saline solution . Two days postinjection, a unilateral stapedotomy was performed . Auditory thresholds were evaluated by electrocochleography before and after stapedotomy . RESULTS: Bacterial OM was induced in all ears injected with pneumococci and pseudomonas . Auditory thresholds increased in all ears treated with stapedotomy . Stapedotomy performed in the presence of pseudomonas OM resulted in significantly greater SNHL than in control or pneumococcal ears (P = 0.0055) . Hearing thresholds were not significantly different between pneumococcal OM and control ears treated with stapedotomy . CONCLUSION: Stapedotomy in the presence of pseudomonas OM, but not pneumococcal OM, significantly increases the risk of SNHL in the guinea pig model . SIGNIFICANCE: Stapedotomy should be avoided in patients with active OM, particularly in the presence of pseudomonas.

J Med Microbiol, 2004 Jan, 53(Pt 1), 61 - 5
Antibacterial activity of the marine sponge constituent cribrostatin 6; Pettit RK et al.; The antibacterial activity of the nitrogen heterocyclic sponge constituent cribrostatin 6 was examined . Cribrostatin 6 was bacteriostatic for a variety of Gram-positive species and was bactericidal for the majority of clinical isolates of Streptococcus pneumoniae, including penicillin-resistant strains . Minimum bactericidal concentration/MIC ratios were < or =2 for 75 % of S . pneumoniae clinical isolates . Kill-curve analysis confirmed the bactericidal action of cribrostatin 6 . Bactericidal activity was rather slow, beginning at 2, 4 or 8 h, depending on the strain . The frequency of occurrence of bacterial spontaneous mutations to resistance was < or =10(-7) . The maximum tolerated dose of cribrostatin 6 in mice was 750-1000 micro g kg(-1) day(-1) . Cribrostatin 6 is a promising lead antibiotic for Gram-positive bacteria, particularly S . pneumoniae, a leading cause of infection and mortality worldwide.

J Med Microbiol, 2004 Jan, 53(Pt 1), 1 - 7
Fate of Streptococcus pyogenes and epithelial cells following internalization; Marouni MJ et al.; The fate of GAS and epithelial cells following internalization was determined in this study . HEp-2 cells harbouring intracellular bacteria were treated with antibiotics to kill extracellular adherent bacteria, washed, and the fate of bacteria and epithelial cells was assessed up to 24 h post-infection . In the absence of antibiotics, massive bacterial growth was apparent in the cell medium, accompanied by extensive cell death, suggesting that intracellular bacteria had multiplied and damaged the monolayer . Addition of the internalization inhibitor, cytochalasin D, either pre- or post-internalization prevented bacterial growth and cell injury; post-internalization treatment with chloramphenicol had the same effect . Analysis of three apoptotic markers in HEp-2 cells - chromatin condensation, DNA laddering and translocation of phosphatidylserine onto the cell-surface membrane - indicated that HEp-2 cells underwent apoptosis . Taken together, the data presented here support a model in which internalized bacteria can induce their own externalization into the medium by a process that requires both an intact host-cell cytoskeleton and de novo synthesis of bacterial proteins . Concomitantly, intracellular and, apparently, extracellular free bacteria induce apoptosis through their cytotoxic activity, and release essential nutrients required for their growth.

J Clin Microbiol, 2003 Dec, 41(12), 5787 - 91
Dissemination of macrolide-resistant Streptococcus pneumoniae isolates containing both erm(B) and mef(A) in South Korea; Waites KB et al.; Macrolide resistance was detected in 64 of 77 (83.1%) Streptococcus pneumoniae isolates from South Korea . Seven (10.9%) isolates contained only mef(A), 32 (50%) contained only erm(B), and 25 (39.1%) contained mef(A) and erm(B) . Nineteen isolates containing mef(A) and erm(B) belonged to serotype 19F, and seven isolates were identical to the Taiwan(19F)-14 clone.

J Clin Microbiol, 2003 Dec, 41(12), 5633 - 9
Molecular epidemiology of penicillin-susceptible non-beta-lactam-resistant Streptococcus pneumoniae isolates from Greek children; Bogaert D et al.; A total of 128 Streptococcus pneumoniae isolates that were susceptible to penicillin but resistant to non-beta-lactam agents were isolated from young carriers in Greece and analyzed by antibiotic susceptibility testing, serotyping, restriction fragment end labeling (RFEL), and antibiotic resistance genotyping . The serotypes 6A/B (49%), 14 (14%), 19A/F (11%), 11A (9%), 23A/F (4%), 15B/C (2%), and 21 (2%) were most prevalent in this collection . Of the isolates, 65% were erythromycin resistant, while the remaining isolates were tetracycline and/or trimethoprim-sulfamethoxazole resistant . Fifty-nine distinct RFEL types were identified . Twenty different RFEL clusters, harboring 2 to 19 strains each, accounted for 76% of all strains . Confirmatory multilocus sequence typing analysis of the genetic clusters showed the presence of three international clones (Tennessee(23F)-4, England(14)-9, and Greece(6B)-22) representing 30% of the isolates . The erm(B) gene was present in 70% of the erythromycin-resistant isolates, whereas 18 and 8% contained the mef(A) and mef(E) genes, respectively . The pneumococci representing erm(B), erm(A), and mef genes belonged to distinct genetic clusters . In total, 45% of all isolates were tetracycline resistant . Ninety-six percent of these isolates contained the tet(M) gene . In conclusion, penicillin-susceptible pneumococci resistant to non-beta-lactams are a genetically heterogeneous group displaying a variety of genotypes, resistance markers, and serotypes . This suggests that multiple genetic events lead to non-beta-lactam-resistant pneumococci in Greece . Importantly, most of these genotypes are capable of disseminating within the community.

J Clin Microbiol, 2003 Dec, 41(12), 5582 - 7
Comparison of antibiotic resistance and serotype composition of carriage and invasive pneumococci among Bangladeshi children: implications for treatment policy and vaccine formulation; Saha SK et al.; The nasopharyngeal carriage of Streptococcus pneumoniae is thought to pose a risk for invasive pneumococcal diseases, and the evaluation of carriage strains is thus often used to inform antibiotic treatment and vaccination strategies for these diseases . In this study, the age-specific prevalences, resistance to antibiotics, and serotype distributions of 1,340 carriage strains were analyzed and compared to 71 pneumococcal strains isolated from the cerebrospinal fluid of children under 5 years old with meningitis . Overall, the nasal carriage rate was 47% . One-fourth (26%) of the infants under 1 month of age and one-half (48%) of the infants under 12 months of age were colonized with S . pneumoniae . Rural children were colonized earlier than those from urban areas . Approximately one-fourth and one-half of the cases of pneumococcal meningitis occurred in the first 3 and 6 months of life, respectively . The respective rates of resistance for carriage and meningitis strains to penicillin (7 and 3%), cotrimoxazole (77 and 69%), and erythromycin (2 and 1%) were similar, whereas chloramphenicol resistance was lower among carriage strains (3%) than among meningitis strains (15.5%) . The predominant serogroups of carriage and invasive isolates were variable and widely divergent . Thus, hypothetical 7-, 9-, and 11-valent vaccines, based on the predominant carriage strains of the present study, would cover only 23, 26, and 30%, respectively, of the serotypes causing meningitis . Further, currently available 7-, 9-, and 11-valent vaccines would protect against only 26, 43, and 48%, respectively, of these meningitis cases . In conclusion, while the surveillance of carriage strains for resistance to antibiotics appears useful in the design of empirical treatment guidelines for invasive pneumococcal disease, data on the serotypes of carriage strains have limited value in vaccine formulation strategies, particularly for meningitis cases.

J Clin Microbiol, 2003 Dec, 41(12), 5541 - 5
Antibiotic-resistant invasive pediatric Streptococcus pneumoniae clones in Israel; Greenberg D et al.; Antibiotic-resistant international clones of Streptococcus pneumoniae are increasingly reported in different parts of the world . We investigated the spread of these clones through an active surveillance performed at the Israeli Streptococcal National Center during 1998 and 1999 . Isolates were tested for antibiotic susceptibility, serotyped, and genotyped by random amplified polymorphic DNA analysis and pulsed-field gel electrophoresis . Of 437 isolates, 276 (63.4%) were antibiotic resistant and 156 (35%) were penicillin nonsusceptible (PNS) . The PNS isolates were less frequently encountered in southern Israel (27 of 136 {20%}) than in other regions (127 of 301 {42%}) . Among 276 antibiotic-resistant isolates, 43 fingerprint patterns were observed . The most common clones were 9V/14-a (19.2%), 5-a (17.8%), and 1-a (10%) . The 9V/14-a clone was less common, while the 1-a clone was more frequent in the south than in other regions . The 5-a clone was more common in Jerusalem than in other regions . Among the Jewish and Arab populations the most frequent clones were 9V/14-a (20%) and 1-a (25%), respectively . Three international clones, 9V/14-a-Spain(9V)-3, 6B-a-Spain(6B)-2, and 5-a-Colombia(5)-19, comprised 40% of all antibiotic-resistant isolates and 56% of all PNS isolates . The seven-valent conjugate vaccine covers 58% of the most common clones, all highly PNS clones, and 94% of the multidrug-resistant clones in Israel, while the nine-valent vaccine covers all of them . The most common antibiotic-resistant invasive pediatric S . pneumoniae clones-mainly the three international ones-contribute significantly to increases in antibiotic resistance . Their geographic distribution varies within the country and between the different populations.

J Clin Microbiol, 2003 Dec, 41(12), 5398 - 406
Evolution of sfbI encoding streptococcal fibronectin-binding protein I: horizontal genetic transfer and gene mosaic structure; Towers RJ et al.; Streptococcal fibronectin-binding protein is an important virulence factor involved in colonization and invasion of epithelial cells and tissues by Streptococcus pyogenes . In order to investigate the mechanisms involved in the evolution of sfbI, the sfbI genes from 54 strains were sequenced . Thirty-four distinct alleles were identified . Three principal mechanisms appear to have been involved in the evolution of sfbI . The amino-terminal aromatic amino acid-rich domain is the most variable region and is apparently generated by intergenic recombination of horizontally acquired DNA cassettes, resulting in a genetic mosaic in this region . Two distinct and divergent sequence types that shared only 61 to 70% identity were identified in the central proline-rich region, while variation at the 3' end of the gene is due to deletion or duplication of defined repeat units . Potential antigenic and functional variabilities in SfbI imply significant selective pressure in vivo with direct implications for the microbial pathogenesis of S . pyogenes.

Obstet Gynecol, 2003 Dec, 102(6), 1332 - 5
The use of a continuous infusion of epinephrine for anaphylactic shock during labor; Gei AF et al.; BACKGROUND: Anaphylaxis is a potentially life threatening, acute, and severe systemic reaction that occurs after the reexposure to a specific antigen . This immunoglobulin E-mediated process is the result of the action of basophils and mast cell mediators, causing severe brochospasm, laringospasm, angioedema, urticaria, and cardiovascular collapse . CASE: We present a case of anaphylactic shock during labor secondary to administration of ampicillin for group B streptococcus prophylaxis . Generalized itching and hives were soon followed by severe maternal hypotension and tachycardia and prolonged fetal bradycardia . These symptoms responded partially to the administration of fluids and parenteral epinephrine . A continuous infusion of epinephrine was required for persistent maternal symptoms . The infusion did not result in further fetal compromise . The patient delivered a healthy fetus 4 hours after the start of the epinephrine infusion . CONCLUSION: This case supports the use of parenteral (intravenous) epinephrine for the treatment of anaphylactic reactions during pregnancy.

Int J Pediatr Otorhinolaryngol, 2003 Dec, 67 Suppl 1, S85 - 90
HIV infection in children--impact upon ENT doctors; Hoare S; The global epidemic of HIV infection remains appalling . By 2001, there were an estimated 1.4 million HIV-infected children, with 4.5 million deaths . In the UK, paediatric cases are clustered around population centres where there are high concentrations of infected immigrant adults, and to a lesser extent, areas where IV drug abuse is common . The highest incidence remains in London and the southeast . With the national redistribution of immigrant and refugee families, any doctor in any specialty may expect to be involved with children who are HIV positive, or have clinical AIDS . The majority of children are infected vertically, i.e . infection of the infant from an infected mother in the pre-, peri-, or post-natal periods . Rates of transmission vary from 15-20% in the developed countries . Children with HIV infection may have their primary presentation to ENT doctors, who should have appropriate thresholds for suspecting the diagnosis . The most common presenting features include persistent generalised lymphadenopathy, hepatosplenomegaly, chronic/recurrent diarrhoea, poor growth, and fever . Fifteen to twenty percent of untreated children will present with an AIDS-defining illness by 12 months, typically with Pneumocystis pneumonia at approximately 3-4 months of age . Seventy percent of perinatally infected children will exhibit some signs or symptoms by 12 months Without treatment, the median age to progression to AIDS is approximately 6 years, and 25-30% will have died by this age . The median age of death is approximately 9 years . Children may also present with repeated/unusual ear infections, sinus disease (inc . mastoiditis), tonsillitis, orbital/peri-orbital cellulitis, oral candidiasis, and dental infections . Infections with streptococcus pneumoniae and group A streptococcus are common, and often progress to severe systemic infection with an appreciable mortality . Infections may be due to unusual pathogens such as Pseudomonas, 'typical' and atypical Mycobacteria, Candida, Aspergillus, etc . Fungal infections of the sinuses (inc . Aspergillus and Rhizopus spp.) may be particularly devastating, with rapid spread to involve bone and the central nervous system . Another classical presentation, which may present to ENT doctors, is that of bilateral parotid enlargement, especially in children who are 'slow progressors', many of whom also have Lymphoid Interstitial Pneumonitis (LIP) . A major attitudinal change has occurred due to advances in 3 main areas: (i) the multidisciplinary management of the infected mother (inc . counselling, antenatal screening, elective caesarean section, advising against breast feeding, etc.), (ii) the prevention of vertical transmission, using anti-retroviral therapy to the infected mother during pregnancy, and to the potentially infected infant in the first weeks of life, and (iii) major advances due to the advent of highly active anti-retroviral treatment . With effective use of these measures, transmission rates may be reduced to <2% . None of the measures though, affect a cure, and it will still be many years before the development of effective vaccines . ENT doctors may be referred children already known to be HIV-positive . Knowing how to talk to infected children (and their parents) is full of potential pitfalls, and requires careful forethought . Many infection-control policies have required considerable rethinking due to the AIDS epidemic . This has especially been the case with respect to needle-stick injuries, post-exposure prophylaxis, sterilization and re-use of equipment, and safe approaches to surgery.

Shanghai Kou Qiang Yi Xue, 2003 Feb, 12(1), 21 - 3
{Arbitrary primed polymerase chain reaction for the genotypic identification of Streptococcus sanguis group}; Shi YC et al.; OBJECTIVE: To find out an ideal method used in identification of Streptococcus sanguis group (SSG) strains by arbitrary primed polymerase chain reaction (AP-PCR) . METHODS: AP-PCR was used to distinguish SSG strains by designing 25 bp arbitrary primer 5' AAG AGA GGA GCT AGC TCT TCT TGG A 3' . RESULTS: There were great differences in the main band of DNA polymorphism among SSG species . The similar band can be obtained from the different DNA extractions in the same species . CONCLUSION: AP-PCR may be useful in the identification and classification of SSG species.

Am Heart J, 2003 Dec, 146(6), 1095 - 8
Clinical profile of Streptococcus agalactiae native valve endocarditis; Rollan MJ et al.; BACKGROUND: Streptococcus agalactiae is an unusual pathogen in adults who are not pregnant . S agalactiae endocarditis is a poorly defined entity because it is uncommon; in contrast to other streptococcal endocarditis, it bears a high mortality rate . The aim of this study was to define its clinical, prognostic, and therapeutic profile on the basis of a series of 9 consecutive patients . METHODS: We conducted a prospective and multicenter study of patients with infectious endocarditis in which 310 episodes were included . RESULTS: S agalactiae grew in 9 patients (3%) who had no valve prosthesis . All patients except 1 had underlying diseases, and all patients had serious complications; the most common complications were major emboli, heart failure, and shock . The valve affected was the mitral valve in 4 patients, the aortic valve in 2 patients, both the mitral and aortic valves in 2 patients, and the tricuspid valve in 1 patient . All episodes were on native valves . Vegetations tended to be large (maximal diameter >10 mm in all patients), very mobile, and pedunculated . An abscess was found in 2 patients, and a perforation of the valve developed in 3 patients . Five patients died (mortality rate, 56%), 3 of whom had received antibiotic therapy alone . The 4 patients who survived underwent combined medical-surgical therapy . CONCLUSION: S agalactiae native valve endocarditis is very aggressive, and early surgery should be considered to prevent the destruction of valves and development of serious complications.

J Biol Chem, 2004 Feb 20, 279(8), 6296 - 304 Epub 2003 Dec 03.
Ectodomains 3 and 4 of human polymeric Immunoglobulin receptor (hpIgR) mediate invasion of Streptococcus pneumoniae into the epithelium; Elm C et al.; Streptococcus pneumoniae binds to the ectodomain of the human polymeric Ig receptor (pIgR), also known as secretory component (SC), via a hexapeptide motif in the choline-binding protein SpsA . The SpsA-pIgR interaction mediates adherence and internalization of the human pathogen into epithelial cells . In this study the results of SpsA binding to human, mouse, and chimeric SC strongly supported the human specificity of this unique interaction and suggested that binding sites in the third and fourth Ig-like domain of human SC (D3 and D4, respectively) are involved in SpsA-pIgR complex formation . Binding of SpsA to SC-derived synthetic peptides indicated surface-located potential binding motifs in D3 and D4 . Adherence and uptake of pneumococci or SpsA-coated latex beads depended on the SpsA hexapeptide motif as well as SpsA-binding sites in D3 and D4 of human pIgR . The involvement of D3 and D4 in adherence and invasion was demonstrated by the lack of binding of SpsA-coated latex beads to transfected epithelial cells expressing mutated pIgR . Finally, blocking experiments with chimeric human-mouse SC as well as synthetic peptides indicated the participation of D3 and a key role of D4 in pneumococcal invasion.

Appl Environ Microbiol, 2003 Dec, 69(12), 7364 - 70
Construction of otherwise isogenic serotype 6B, 7F, 14, and 19F capsular variants of Streptococcus pneumoniae strain TIGR4; Trzcinski K et al.; The polysaccharide capsule is the primary virulence factor in Streptococcus pneumoniae . There are at least 90 serotypes of S . pneumoniae, identified based on the immunogenicity of different capsular sugars . The aim of this study was to construct pneumococcal strains that are isogenic except for capsular type . Serotype 4 strain TIGR4 was rendered unencapsulated by recombinational replacement of the capsular polysaccharide synthesis (cps) locus with the bicistronic Janus cassette (C . K . Sung, J . P . Claverys, and D . A . Morrison, Appl . Environ . Microbiol . 67:5190-5196, 2001) . In subsequent transformation with chromosomal DNA, the cassette was replaced by the cps locus derived from a strain of a different serotype, either 6B, 7F, 14, or 19F . To minimize the risk of uncontrolled recombinational replacements in loci other than cps, the TIGRcps::Janus strain was "backcross" transformed three times with chromosomal DNA of subsequently constructed capsular type transformants . Capsular serotypes were confirmed in all new capsule variants by the Quellung reaction . Restriction fragment length polymorphism (RFLP) analysis of the cps locus confirmed the integrity of the cps region transformed into the TIGR strain, and RFLP of the flanking regions confirmed their identities with the corresponding regions of the recipient . Transformants had in vitro growth rates greater than or equal to that of TIGR4 . All four strains were able to colonize C57BL/6 mice (female, 6 weeks old) for at least 7 days when mice were intranasally inoculated with 6 x 10(6) to 8 x 10(6) CFU . The constructed capsular variants of TIGR4 are suitable for use in studies on the role of S . pneumoniae capsular polysaccharide in immunity, colonization, and pathogenesis.

Int J Antimicrob Agents, 2003 Dec, 22(6), 588 - 93
Bactericidal effect of cethromycin (ABT-773) in an immunocompetent murine pneumococcal pneumonia model; Capitano B et al.; The efficacy of cethromycin was assessed against isolates of Streptococcus pneumoniae in the presence of neutrophils . Comparison with data from our previous neutropenic model revealed that the presence of neutrophils enhanced the bacteriostatic and bactericidal effect of cethromycin by an average of two- to four-times, respectively.

Int J Antimicrob Agents, 2003 Dec, 22(6), 579 - 87
Disposition and intracellular levels of moxifloxacin in human THP-1 monocytes in unstimulated and stimulated conditions; Hall IH et al.; Moxifloxacin uptake by human THP-1 monocytes was passive and initially linear and reached equilibrium after approximately 4 h . High intracellular concentrations were achieved and intracellular/extracellular {I/E} ratios were between 1925 and 4575 for the lowest concentration of 0.004 microg/ml at pH 7.4 and 6.9 . The uptake of moxifloxacin was reduced by sodium fluoride, -azide, -cyanide, low temperature and low pH . However, the uptake was not affected by any of the ion channel blockers . Adenosine demonstrated marginal competition with moxifloxacin for uptake suggesting a nucleoside transporter may be involved . The sodium-ATPase pump when blocked, also retarded moxifloxacin uptake at 2 and 4 h . This I/E ratio was high compared with other macrolides and indicateed that the monocyte may contain sufficient moxifloxacin levels to conduct the antibiotic throughout systemic circulation to infection sites . Efflux from THP-monocytes was essentially complete after 2 h indicating no long term sequestering of the antibiotic occurred . Disposition of the antibiotic within the THP-1 monocytes showed large amounts present in the nucleus and cytoplasm in stimulated and unstimulated cells . Increased amounts of the drug were found in the THP-1 monocytes as well as the endoplasmic reticulum and the isolated phagosomes after stimulation by zymogen A, Staphylococcus aureus or Streptococcus pneumoniae.

Paediatr Drugs, 2003, 5(12), 821 - 32
Community-acquired pneumonia in children: issues in optimizing antibacterial treatment; Korppi M; The treatment of community-acquired pneumonia (CAP) in children is empirical, being based on the knowledge of the etiology of CAP at different ages . As a result of currently available methods in everyday clinical practice, a microbe-specific diagnosis is not realistic in the majority of patients . Even the differentiation between viral, 'atypical' bacterial (Mycoplasma pneumoniae or Chlamydia pneumoniae) and 'typical' bacterial (Streptococcus pneumoniae) CAP is often not possible . Moreover, up to one-third of CAP cases seem to be mixed viral-bacterial or dual bacterial infections . Recent serologic studies have confirmed that S . pneumoniae is an important causative agent of CAP at all ages . M . pneumoniae is common from the age of 5 years onwards, and C . pneumoniae is common from the age of 10 years onwards . In addition to age, the etiology and treatment of CAP are dependent on the severity of the disease . Pneumococcal infections are predominant in children treated in hospital, and mycoplasmal infections are predominant in children treated at home.In ambulatory patients with CAP, amoxicillin (or penicillin V {phenoxymethylpenicillin}) is the drug of choice from the age of 4 months to 4 years, and at all ages if S . pneumoniae is the presumptive causative organism . Macrolides, preferably clarithromycin or azithromycin, are the first-line drugs from the age of 5 years onwards . In hospitalized patients who need parenteral therapy for CAP, cefuroxime (or penicillin G {benzylpenicillin}) is the drug of choice . Macrolides should be administered concomitantly if M . pneumoniae or C . pneumoniae infection is suspected . Radiologic findings and C-reactive protein (CRP) levels offer limited help for the selection of antibacterials; alveolar infiltrations and high CRP levels indicate pneumococcal pneumonia, but the lack of these findings does not rule out bacterial CAP . Most guidelines recommend antibacterials for 7-10 days (except azithromycin, which has a recommended treatment duration of 5 days) . If no improvement takes place within 2 days, therapy must be reviewed.

Int J Syst Evol Microbiol, 2003 Nov, 53(Pt 6), 1941 - 6
rDNA sequence analyses of Streptococcus dysgalactiae subsp . equisimilis isolates from pigs; Kawata K et al.; The nucleotide sequences of 16S and 23S rRNA genes (rDNA) were determined for 11 isolates of Streptococcus dysgalactiae subsp . equisimilis from slaughtered pigs with endocarditis, arthritis or lymphadenitis and strain ATCC 35666, designated as a strain of subspecies equisimilis . The sequences of each of the genes were compared phylogenetically with the corresponding sequences of S . dysgalactiae subsp . dysgalactiae ATCC 43078(T) and ATCC 27957, which were also determined in this study . Based on the 16S rDNA analysis, the isolates of S . dysgalactiae subsp . equisimilis were divided into two distinct groups, designated groups 1 and 2 . S . dysgalactiae subsp . equisimilis ATCC 35666 was closely related to the group 2 strains . The S . dysgalactiae subsp . dysgalactiae strains seemed to be associated with the group 1 strains, which was not consistent with the conventional subspecific classification of S . dysgalactiae . In contrast, the 23S rDNA analysis distinguished S . dysgalactiae subsp . dysgalactiae strains from subsp . equisimilis strains . This inconsistency between phylogenies based on 16S and 23S rDNA indicates that 23S rDNA is a more rigid marker for determining the phylogenetic relationships and taxonomic position of these organisms than is 16S rDNA.

Int J Syst Evol Microbiol, 2003 Nov, 53(Pt 6), 1861 - 71
Phylogenetic relationships and genotyping of the genus Streptococcus by sequence determination of the RNase P RNA gene, rnpB; Tapp J et al.; The rnpB gene is universally present in bacterial species and encodes the RNA subunit of endoribonuclease P . In this study, rnpB was sequenced in 50 type strains and 29 additional strains of the genus Streptococcus . Putative secondary-structure models and possible interactions in RNase P RNA molecules are discussed . Phylogenetic relationships were studied and Bayesian, maximum-parsimony and minimum-evolution analyses supported six main clades that comprised 22 of the 50 species analysed . Phylogenetic inference was also studied for the 16S rRNA gene; it indicated a similar tree topology, but with weaker support values than for rnpB . Combined analysis of rnpB and 16S resulted in a phylogeny with significantly better support . Variability in the rnpB and 16S genes among all type strains, calculated as Shannon-Wiener information index values, was 0.45 for rnpB and 0.15 for 16S . Intraspecies proximity was assessed by principal coordinate analysis of rnpB for 32 strains of six closely related species (two clades) and showed species-specific clusters, but heterogeneity occurred in two species . It can be concluded that the rnpB gene is suitable for phylogenetic analysis of closely related taxa and has potential as a tool for species discrimination.

Structure (Camb), 2003 Dec, 11(12), 1499 - 511
The structure of chorismate synthase reveals a novel flavin binding site fundamental to a unique chemical reaction; Maclean J et al.; The crystal structure of chorismate synthase (CS) from Streptococcus pneumoniae has been solved to 2.0 A resolution in the presence of flavin mononucleotide (FMN) and the substrate 5-enolpyruvyl-3-shikimate phosphate (EPSP) . CS catalyses the final step of the shikimate pathway and is a potential therapeutic target for the rational design of novel antibacterials, antifungals, antiprotozoals, and herbicides . CS is a tetramer with the monomer possessing a novel beta-alpha-beta fold . The interactions between the enzyme, cofactor, and substrate reveal the structural reasons underlying the unique catalytic mechanism and identify the amino acids involved . This structure provides the essential initial information necessary for the generation of novel anti-infective compounds by a structure-guided medicinal chemistry approach.

MMWR Morb Mortal Wkly Rep, 2003 Dec 5, 52(48), 1174 - 6
Invasive Streptococcus pyogenes after allograft implantation--Colorado, 2003; Centers for Disease Control and Prevention (CDC); Allograft tissues are used for various orthopedic procedures (e.g., ligament reconstruction, meniscal transplantation, and spinal surgery) . In 2002, approximately one million allografts were distributed for transplantation (American Association of Tissue Banks {AATB}, unpublished data, 2002) . Recent reports of allograft-associated infections have prompted evaluation of the processing and quality-control methods employed by tissue processors . This report describes a case of invasive disease with Streptococcus pyogenes (i.e., group A streptococcus {GAS}), after reconstructive knee surgery using contaminated allograft tissue and provides recommendations to reduce the risk for allograft-associated infections . Although allograft infections are rare, they highlight the need for improved tissue evaluation and processing standards.

Eur J Clin Microbiol Infect Dis, 2004 Jan, 23(1), 27 - 33 Epub 2003 Dec 02.
Multicentre study of the molecular epidemiology, serotypes and antimicrobial susceptibility patterns of invasive Streptococcus pneumoniae invasive isolated from children in the Ille de France area; Decousser JW et al.; Between January 1997 and April 2002, 73 consecutive invasive strains of Streptococcus pneumoniae were isolated from children under 16 years of age in four hospitals in suburban Paris . Their genetic diversity was investigated by serotyping and analysis of pulsed-field gel electrophoresis restriction patterns . Antibiotic susceptibility patterns were analysed by disk susceptibility testing and determination of minimal inhibitory concentrations . The genetic basis of macrolide resistance was investigated by polymerase chain reaction . Studies of penicillin and vancomycin tolerance were performed for each strain . Despite the high prevalence (45.2%) of penicillin-nonsusceptible Streptococcus pneumoniae, resistance to amoxicillin (1.4%) was rare, and no strain was resistant to cefotaxime . Overall, 4.1% of pneumococcal strains were resistant to penicillin . Penicillin or vancomycin tolerance was not detected in any of the 73 strains studied . Of the erythromycin-resistant strains (48%), all but one carried the ermB gene . No strains showing a decreased susceptibility to ciprofloxacin (MIC, >4 mg/l) or overexpressing an efflux pump inhibited by reserpine were isolated . The serotypes found, in order of frequency, were as follows: 18C, 14, 6B, 19F, 19A, 9V, 23F, 1, 7F, 9A, 38 . Strains of penicillin-nonsusceptible Streptococcus pneumoniae belonged predominantly to serotypes 14, 6B, 9V, 9A, 23F, 19F and 19A . The seven-valent conjugated vaccine covered 85.5% of the serogroups isolated in children under 2 years of age and 65.6% of the serogroups identified in children over 2 years of age . The genetic analysis showed a high identity for some serotypes, such as 14/9V, 6B and 23F . The use of the seven-valent conjugated vaccine is a critical measure to prevent invasive pneumococci infections in children in the Ille de France area.

Mol Microbiol, 2003 Dec, 50(5), 1647 - 63
Constitutive expression of PcsB suppresses the requirement for the essential VicR (YycF) response regulator in Streptococcus pneumoniae R6; Ng WL et al.; We report several new findings about the function of the essential VicRK two-component regulatory system (TCS) in the human pathogen Streptococcus pneumoniae . The vicR-encoded response regulator, vicK-encoded histidine kinase and the protein encoded by the downstream vicX gene are the homologues of the YycF, YycG and YycJ proteins, respectively, studied previously in Bacillus subtilis and Staphylococcus aureus . Using a regulatable promoter, we demonstrated that the VicK histidine kinase is conditionally required for growth of S . pneumoniae . Likewise, we found that the VicX protein is also conditionally required for growth and probably plays a role in the essential signal transduction pathway mediated by VicR and VicK . Recovery of limited substitutions in the conserved aspartate 52 residue (D52) of VicR was consistent with a requirement for phosphorylation of VicR for growth under some conditions . We applied microarrays to characterize the changes in transcription patterns in bacteria depleted for vicRKX operon expression . Our results suggest that the pcsB gene is a target of the VicRK TCS . We present evidence that downregulation of pcsB could account for many of the defects in cell growth, shape, size and morphology observed in bacteria depleted for vicRKX expression . Furthermore, constitutive expression of pcsB+ suppressed the essential requirement for the VicRK TCS and allowed the isolation of vicR null mutants.

Mol Microbiol, 2004 Jan, 51(1), 123 - 34
Invasive M1T1 group A Streptococcus undergoes a phase-shift in vivo to prevent proteolytic degradation of multiple virulence factors by SpeB; Aziz RK et al.; A globally disseminated strain of M1T1 group A Streptococcus (GAS) has been associated with severe infections in humans including necrotizing fasciitis and toxic shock syndrome . Recent clinicoepidemiologic data showed a striking inverse relationship between disease severity and the degree to which M1T1 GAS express the streptococcal cysteine protease, SpeB . Electrophoretic 2-D gel analysis of the secreted M1T1 proteome, coupled with MALDI-TOF mass spectroscopy, revealed that expression of active SpeB caused the degradation of the vast majority of secreted GAS proteins, including several known virulence factors . Injection of a SpeB+/SpeA- M1T1 GAS strain into a murine subcutanous chamber model of infection selected for a stable phase-shift to a SpeB-/SpeA+ phenotype that expressed a full repertoire of secreted proteins and possessed enhanced lymphocyte-stimulating capacity . The proteome of the SpeB-in vivo phase-shift form closely matched the proteome of an isogenic speB gene deletion mutant of the original M1T1 isolate . The absence or the inactivation of SpeB allowed proteomic identification of proteins in this M1T1 clone that are not present in the previously sequenced M1 genome including SpeA and another bacteriophage-encoded novel streptodornase allele . Further proteomic analysis of the M1T1 SpeB+ and SpeB- phase-shift forms in the presence of a cysteine protease inhibitor demonstrated differences in the expression of several proteins, including the in vivo upregulation of SpeA, which occurred independently of SpeB inactivation.

Pediatr Dermatol, 2003 Nov-Dec, 20(6), 506 - 10
Childhood pustular psoriasis elicited by the streptococcal antigen: a case report and review of the literature; Cassandra M et al.; The Zumbusch pattern of generalized pustular psoriasis (GPP) classically presents as waves of widespread sheets of sterile pustules on brightly erythematous skin . The occurrence of this disease in childhood is rare, and fewer than 200 cases have been reported in the literature . We describe a 10-year-old boy with GPP who had an elevated serum antistreptolysin titer . Several antigenic factors shown to elicit GPP have been reported, including withdrawal of steroids, emotional stress, and infection . However, we further propose that the group A beta-hemolytic streptococcus can trigger a flare of GPP . We suggest that if pustular psoriasis is suspected clinically, an elevated serum antistreptolysin antibody titer may help identify the causative antigen.

Hua Xi Kou Qiang Yi Xue Za Zhi, 2003 Oct, 21(5), 396 - 9
{An experimental study on PAc and GTF gene vaccines of Streptococcus mutans against rats caries: antibody levels in saliva and serum}; Yang D et al.; OBJECTIVE: The purpose of this study is to examine the levels of salivary SIgA and serum IgG induced by pcDNA3-pac and pcDNA3-gtfB immunization, so as to testify the antigenity of the two gene vaccines . METHODS: 36 28-day-old Wistar rats were divided into 6 groups, among which 3 experimental groups were vaccinated with pcDNA3-pac, pcDNA3-gtfB or pcDNA3-pac combined with pcDNA3-gtfB, respectively, one positive control was vaccinated with inactive whole cell of S . mutans JBP and other two negative controls were injected with the vector pcDNA3 or PBS buffer, respectively . All vaccines and materials were delivered with 100 micrograms by submandibular gland injection for 3 times . Then the restricted bacterial model of rat was constructed . Following that all rats were fed with cariogenic diet Keyes 2000 for 3 months, saliva and serum samples were collected to assay SIgA or IgG levels by ELASA . RESULTS: The salivary S-IgA levels both in pcDNA3-pac combined with pcDNA3-gtfB group and inactive S . mutans cell group were higher than others (P < 0.01) . In groups of pcDNA3 and PBS buffer, they were lowest (P < 0.01) . The serum IgG levels in the three experimental groups and positive control were higher than that in negative control (P < 0.05) . It was important that salivary SIgA in groups of gene vaccine and inactive S . mutans vaccination reached its peak at the 11th week after the first inoculation and kept until the end of the study . CONCLUSION: Both pcDNA3-pac and pcDNA3-gtfB can express immunogenic protein and induce immune responses of mucosal and humoral immune system in gnobobiotic rats . It is also indicated that the joint gene vaccines immunization is an optimal choice for anticaries strategy.

Hua Xi Kou Qiang Yi Xue Za Zhi, 2003 Oct, 21(5), 353 - 5
{The study of peptide vaccine HDS from Streptococcus mutans glucosyltransferase}; Ding Y et al.; OBJECTIVE: To investigate the antigenicity of the peptide vaccine HDS from Streptococcus mutans glucosyltransferase and its ability to induce protective immune responses in an experimental rat model of dental caries . METHODS: Artificial antigen HDS-KLH, peptide HDS, glycosyltransferase were injected to immunize rats . Measurement of the specific anti-HDS, GTF IgG or IgA concentration in saliva and serum were undertaken by ELISA among the experimental groups . Gnotobiotic rat model was developed when challenged S . mutans and a caries promoting diet . The jaws of the rats were selected and dyed . The Keyes caries score for each jaw were counted . RESULTS: The level of serum and salivary specific anti-HDS IgG and IgA in the group immunized by HDS-KLH was significantly higher than that in control group (P < 0.05) . The Keyes caries score of GTF, HDS and HDS-KLH immunized group were significantly lower than that of control group, especially lower in smooth tooth surface . CONCLUSION: Artificial antigen HDS-KLH could induce immune response . As a peptide vaccine, HDS-KLH could reduce the caries incidence in experimental rat model.

Hua Xi Kou Qiang Yi Xue Za Zhi, 2003 Oct, 21(5), 350 - 2
{DNA sequence analysis for the promoter of pyruvate oxidase gene from Streptococcus oralis}; Zhang J et al.; OBJECTIVE: To elucidate the molecular structure of pyruvate oxidase gene promoter . METHODS: The 1.30 kb fragment with promoter activity, amplified from upstream of Streptococcus oralis pyruvate oxidase gene (Sopox), was cloned into vector PBK-CMV . The positive transformed E . coli JM109 clone was selected, the recombinant plasmid was further identified with restriction mapping analysis . The positive recombinant plasmid was studied with sequence analysis . RESULTS: After digesting the recombinant plasmid with Hind III, 1% agarose electrophoresis showed 1.30 kb fragment, which was consistent with predicted size . Sequence analysis revealed 1,350 bp . CONCLUSION: The Sopox promoter region is sequenced . Further characterization of the Sopox promoter region will elucidate the molecular mechanism of H2O2 production of streptococcus oralis.

Hua Xi Kou Qiang Yi Xue Za Zhi, 2003 Oct, 21(5), 339 - 41
{The study on specific anti-Streptococcus mutans IgY against dental caries in rats}; Fan M et al.; OBJECTIVE: To investigate the effect of specific anti-streptococcus mutans IgY against streptococcus mutans on dental caries development in rats . METHODS: 35 wistar rats were divided into 5 groups: group A received IgY gargle; group B received IgY lyophilized powder; group C received sterilized water as control; group D and E received egg yolk food with or without specific IgY individually . They were all fed with caries-inducing diet 2000# . The number of caries scores was counted by the procedure of Keyes' . RESULTS: There was a significant lower mean of caries scores in groups treated with IgY lyophilized powder and gargle . By treating with egg-yolk food contained specific IgY, the mean of caries scores decreased comparing with no treatment group . CONCLUSION: Local passive immunization with specific anti-streptococcus mutans IgY may be an effective way to prevent the development of dental caries.

Arch Gynecol Obstet, 2004 Jan, 269(2), 139 - 41 Epub 2003 Jan 11.
Maternal postpartum group B beta-hemolytic streptococcus ventriculoperitoneal shunt infection; Kane JM et al.; BACKGROUND: Women with cerebrospinal fluid shunts require special management during the course of pregnancy . CASE REPORT: We describe a case of delayed postpartum ventriculoperitoneal shunt infection by Group B streptococcus in a 19-year-old who presented complaining of headache and a fever . The CSF culture from the shunt tap and the distal shunt tip both grew Group B beta-hemolytic streptococcus . CONCLUSION: Women who are colonized with Group B streptococcus and who have cerebrospinal fluid shunts should receive perinatal antibiotic prophylaxis, and may require more extended prophylactic antibiotics with cesarean section deliveries to prevent catheter tip colonization and subsequent shunt infection.

HNO, 2003 Dec, 51(12), 986 - 92
{Therapeutic management of necrotizing neck infections}; Rudack C et al.; BACKGROUND: Necrotizing neck infections are uncommon soft-tissue infections, usually caused by virulent, toxin producing bacteria . Necrotizing fasciitis represents a special form of necrotizing soft tissue infection with a mortality rate of up to 76% even though aggressive therapy is recommended . PATIENTS AND METHODS: In the last 2 years we treated four patients with severe necrotizing neck infections and five suffering from necrotizing fasciitis . RESULTS: Microbiological analysis revealed mixed infections with Candida albicans, Streptococcus pyogenes, Fusobacterium, Proprioni bacteria and Staphylococcus . The surgical management was not only restricted to drainage, but also included functional neck dissection in order minimize the spread of the disease . Eight of our patients recovered completely, but one died due to toxic shock as consequence of a delayed in therapy . CONCLUSION: Complete recovery of patients suffering from necrotizing fasciitis depends on early and aggressive surgical therapy including neck dissection and drainage as well as an interdisciplinary strategy of conservative therapy . Hyperbaric oxygen should be considered as a treatment adjunct in patients with necrotizing fasciitis if surgery and antibiotic treatment fail.

J Pediatr (Rio J), 2002 Jan-Feb, 78(1), 19 - 23
{Pneumococcal meningitis in children: clinical findings, most frequent serotypes and outcome}; Berezin EN et al.; OBJECTIVE: To determine mortality, morbidity, antimicrobial susceptibility and the most frequent serotypes in children admitted to hospital due to pneumococcal meningitis METHODS: Patients with meningitis caused by Streptococcus pneumoniae detected by culture in cerebrospinal fluid or blood, aged between 1 month and 15 years old, admitted to two hospitals in the city of Sao Paulo, were included in the study . Susceptibility to penicillin was determined by the disk diffusion test using oxacillin 1 micro g disk . If the inhibition area with oxacillin disk was less than 20mm, the strains were tested for penicillin, chloramphenicol, ceftriaxone, vancomycin and sulfamethoxazole /trimethoprim using the E test . RESULTS: We identified 55 patients, 52.5% of which were younger than 6 months . The prevalence of penicillin-nonsusceptible strains was 36% . All the strains were intermediately resistant (0.1 micro g/ ml </=; MIC</= 1.0 micro g/ ml) and 35% of the penicillin intermediate resistant strains were resistant to sulfamethoxazole/trimethoprim . The mortality rate was 20% and impaired neurological outcome was present in 40% of the children . The audiometric test revealed alteration in 60% of the children tested . Age less than 6 months was associated with poor outcome . The most frequent serotypes were 1, 5, 6B, 14, 19A and 23F, and 70% of the serotypes were included in the new 7-valent vaccine . CONCLUSIONS: These findings suggest that pneumococcal meningitis presents high mortality and morbidity and that the 7-valent conjugate vaccine would be potentially useful in preventing serious pneumococcal infections.

J Pediatr (Rio J), 2001 May-Jun, 77(3), 227 - 34
{Nasopharyngeal colonization and antimicrobial resistance of Streptococcus pneumoniae isolated from children with acute rhinopharyngitis}; Ferreira LL et al.; OBJECTIVE: To determine the prevalence and risk factors for nasopharyngeal colonization by, and to evaluate antimicrobial susceptibility of Streptococcus pneumoniae strains in children with acute rhinopharyngitis . METHODS: We collected nasopharyngeal swab specimens from 400 children aged 3 months to 5 years and with clinical status of acute rhinopharyngitis from June 16, 1997 to May 20, 1998 at the outpatient clinics of two hospitals in the city of Sao Paulo . Nasopharyngeal specimens were collected pernasally using a calcium alginate swab and plated immediately after collection onto trypticose soy agar with 5% sheep blood and garamicin 5 mcg/ml . Penicillin susceptibility was determined by oxacillin 1 mcg disk screening test and the minimal inhibitory concentration by the E-test . RESULTS: Pneumococci were recovered from 139 children, indicating a colonization prevalence of 35% . The risk factors analyzed indicated that the colonization was more prevalent in children attending day-care centers, children with siblings younger than 5 years, and children with recent use of antimicrobial agents . The prevalence of penicillin non-susceptible strains was of 16 % (20 strains) . All strains were intermediately resistant (0.1mcg/ ml </= MIC </= 1.0 mcg/ ml) . Out of the penicillin intermediately resistant strains, 7 (37%) showed intermediate resistance to cotrimoxazol and 2 (11%) full resistance to trimethoprim-sulfamethoxazole . No strains were resistant to ceftriaxone, amoxicillin, clarithromicin, or chloramphenicol . CONCLUSIONS: Our findings indicate that the prevalence of nasopharyngeal colonization by Streptococcus pneumoniae in children with upper respiratory infections was of 34.8% . Children attending day-care centers and children with younger siblings showed higher levels of colonization The results of prevalence of bacterial resistance were similar to those of studies regarding invasive infections, thus indicating that nasopharyngeal isolates of Streptococcus pneumoniae can be used in the surveillance of antimicrobial resistance in a defined geographical area.

Saudi Med J, 2003 Nov, 24(11), 1210 - 3
Penicillin resistance in serogroups/serotypes of Streptococcus pneumoniae causing invasive infections in Central Saudi Arabia; Twum-Danso K et al.; OBJECTIVE: To determine the minimum inhibitory concentrations (MICs) of penicillin, ceftriaxone and vancomycin of serogroups/serotypes of Streptococcus pneumoniae (S . pneumoniae) from invasive diseases in all age groups from major hospitals in Riyadh, Kingdom of Saudi Arabia (KSA) . METHODS: All isolates of S . pneumoniae from patients with invasive pneumococcal infections between February 2000 and November 2001 were prospectively collected from 8 major hospitals in Riyadh, KSA . The isolates were confirmed as S . pneumoniae at the King Khalid University Hospitals, Riyadh, KSA and then serogrouped/serotyped using the agglutination method . The MICs for penicillin, ceftriaxone and vancomycin were carried out using the E-test . RESULTS: Forty-three percent of the isolates were resistant to penicillin mostly of the intermediate type (97%) . The resistant strains were mainly confined to serogroups/serotypes 6, 23, 19 and 15 and the 7-valent conjugate vaccine covers 76% of the penicillin-resistant strains . Only one isolate was resistant to ceftriaxone . CONCLUSION: In view of the rather insignificant level of highly resistant-penicillin strains and the virtual absence of resistance to ceftriaxone we would like to suggest using ceftriaxone for treating invasive pneumococcal infections outside the central nervous system . We recommend that the conjugate vaccine would be a useful adjunct to penicillin prophylaxis in patients at risk in our community.

J Bacteriol, 2003 Dec, 185(24), 7241 - 6
Horizontal transfer of segments of the 16S rRNA genes between species of the Streptococcus anginosus group; Schouls LM et al.; The nature in variation of the 16S rRNA gene of members of the Streptococcus anginosus group was investigated by hybridization and DNA sequencing . A collection of 708 strains was analyzed by reverse line blot hybridization . This revealed the presence of distinct reaction patterns representing 11 different hybridization groups . The 16S rRNA genes of two strains of each hybridization group were sequenced to near-completion, and the sequence data confirmed the reverse line blot hybridization results . Closer inspection of the sequences revealed mosaic-like structures, strongly suggesting horizontal transfer of segments of the 16S rRNA gene between different species belonging to the Streptococcus anginosus group . Southern blot hybridization further showed that within a single strain all copies of the 16S rRNA gene had the same composition, indicating that the apparent mosaic structures were not PCR-induced artifacts . These findings indicate that the highly conserved rRNA genes are also subject to recombination and that these events may be fixed in the population . Such recombination may lead to the construction of incorrect phylogenetic trees based on the 16S rRNA genes.

J Bacteriol, 2003 Dec, 185(24), 7176 - 83
Competence-induced cells of Streptococcus pneumoniae lyse competence-deficient cells of the same strain during cocultivation; Steinmoen H et al.; Several streptococcal species are able to take up naked DNA from the environment and integrate it into their genomes by homologous recombination . This process is called natural transformation . In Streptococcus pneumoniae and related streptococcal species, competence for natural transformation is induced by a peptide pheromone through a quorum-sensing mechanism . Recently we showed that induction of the competent state initiates lysis and release of DNA from a subfraction of the bacterial population and that the efficiency of this process is influenced by cell density . Here we have further investigated the nature of this cell density-dependent release mechanism . Interestingly, we found that competence-induced pneumococci lysed competence-deficient cells of the same strain during cocultivation and that the efficiency of this heterolysis increased as the ratio of competent to noncompetent cells increased . Furthermore, our results indicate that the lysins made by competent pneumococci are not released into the growth medium . More likely, they are anchored to the surface of the competent cells by choline-binding domains and cause lysis of noncompetent pneumococci through cell-to-cell contact.

Arch Pediatr, 2003 Dec, 10(12), 1056 - 60
{Community acquired pneumonia and influenza in children}; Pons-Catalano C et al.; Children without chronic or serious medical conditions are at increased risk for hospitalization during influenza seasons, mainly with respiratory tract infections . But influenza virus infections frequently remain undiagnosed, even in hospitalized patients . We prospectively studied the rate of concomitant and preceding influenza infections in children hospitalized with a community acquired pneumonia (CAP) . POPULATION AND METHODS: All 1-15-year-old children with CAP requiring hospitalization between 1st April 2000 and 2002 had nasopharyngeal aspirate for viruses, immunofluorescence and serologies for respiratory pathogens . The peak of influenza IgG measured by complement fixation (CF) is transient, and a titer of 1/64 or more indicates an acute influenza infection in the preceding weeks . Children with chronic disease were excluded and a control group of patients from outpatient clinic was measured . RESULTS: Among 33 previously healthy children (age 4.9 years, range 1.2-14 years), 8 had a pneumococcal pneumonia, 10 a pneumonia caused by Mycoplasma pneumoniae (MP), 1 by Chlamydia pneumonia, and 8 of unknown origin . In six patients immunofluorescence was positive: Respiratory Syncitial Virus, 2, Adenovirus, 1 and Influenza A, 3 (including a patient with concomitant MP infection) . Thirteen of the 33 children (39.4%) had evidence of a recent influenza A infection with CF ab > or = 1/64: with pneumococcal pneumonia, 5/10 with MP pneumonia, 3/8 with unknown origin pneumonia, 9/13 of these previous influenza infections being clinically inapparent . Only 1/30 children of control group (3.3%) had CF ab > or = 1/64 . CONCLUSION: In this study, influenza infection is the direct cause of CAP of children in 12% of cases . In other children with CAP, 39.4% of patients had an influenza infection in the preceding weeks which leads to secondary infection caused by Streptococcus pneumoniae or by MP or other pathogens.

FEMS Immunol Med Microbiol, 2003 Dec 5, 39(3), 265 - 73
Epitope mapping of pneumococcal surface protein A of strain Rx1 using monoclonal antibodies and molecular structure modelling; Kolberg J et al.; Pneumococcal surface protein A (PspA) is an antigenic variable vaccine candidate of Streptococcus pneumoniae . Epitope similarities between PspA from the American vaccine candidate strain Rx1 and Norwegian clinical isolates were studied using PspA specific monoclonal antibodies (mAbs) made against clinical Norwegian strains . Using recombinant PspA/Rx1 fragments and immunoblotting the epitopes for mAbs were mapped to two regions of amino acids, 1-67 and 67-236 . The discovered epitopes were visualized by modelling of the PspA:Fab part of mAb in three dimensions . Flow cytometric analysis showed that the epitopes for majority of mAbs were accessible for antibody binding on live pneumococci . Also, the epitopes for majority of the mAbs are widely expressed among clinical Norwegian isolates.

Mycoses, 2003 Dec, 46(11-12), 471 - 8
In vitro cariogenic potential of Candida albicans; Nikawa H et al.; The adherence and dissociation of Candida albicans, C . tropicalis, Streptococcus mutans and S . sanguis to six substrates including hydroxylapatite (HAP) which exhibit various hydrophobicity, was examined by the use of a bioluminescent adenosine triphosphate (ATP) assay . Dissolution of HAP by C . albicans or S . mutans was determined spectrophotometrically by the use of o-cresolphthalein complexone . In the adherence of C . tropicalis, S . mutans and S . sanguis, the amount of adherent cells correlated with the hydrophobicity of the substrates . In contrast, the adherence of C . albicans to HAP was extraordinary high, although the adherence of the fungi also correlated with the hydrophobicity of the substrates, except for HAP . The yeasts attached to HAP was effectively removed by high concentration of either phosphate or calcium ions . The amount of calcium-release from HAP caused by C . albicans and S . mutans was 113 microg ml(-1) (final pH = 3.45), and 5.4 microg ml(-1) (final pH 4.81), respectively and the maximum growth of C . albicans and S . mutans was 10(7) cfu ml(-1) and 7.4 x 10(12) cfu ml(-1), respectively . The results, taken together, suggest that C . albicans adhere to HAP specifically through electrostatic interaction, and that, in a much smaller number (1.0/7.4 x 10(5)), C . albicans possesses the ability to dissolve HAP to a greater extent (approximately 20-fold) when compared with S . mutans.

Jpn J Thorac Cardiovasc Surg, 2003 Nov, 51(11), 616 - 8
Cusp commissuroplasty for tricuspid valve endocarditis; Hamanaka Y et al.; A 19-year-old woman with a history of drug addiction suffered from sepsis and heart failure . Blood culture was positive for Streptococcus viridans . An operation was indicated because the echocardiography showed massive vegetation on the anterior leaflet of the tricuspid valve and severe regurgitation even though the endocarditis was healed with drug therapy . At operation all of the anterior leaflet of the tricuspid valve was resected with the vegetation . Using the technique of cusp commissuroplasty, the disrupted commissure was reconstructed by approximating the septal and posterior cusps at the level of their normal closure, forming a zone of apposition by using a single stitch . Leaflet apposition resulted in a defect between the apposed leaflets and the tricuspid annulus, which was patched with autologous pericardium . The tricuspid valve was reconstructed to function as a unicommissural bicuspid valve . The patient was stable during the follow-up period of two years without any medical treatment.

Postgrad Med, 2003 Nov, 114(5), 43 - 50; quiz 8
New issues in bacterial meningitis in adults . Antibiotic resistance has complicated treatment; Spach DH; The epidemiologic factors of bacterial meningitis, a serious disease that must be addressed with great urgency, have evolved dramatically in the last 25 years . Among both adults and children, multidrug-resistant Streptococcus pneumoniae has emerged as a clinical challenge and has greatly complicated the empirical management of this disease . In this article, the author focuses on new issues involving the epidemiologic factors, diagnosis, treatment, and prevention of bacterial meningitis in adults.

Infect Control Hosp Epidemiol, 2003 Nov, 24(11), 848 - 52
A preventable outbreak of pneumococcal pneumonia among unvaccinated nursing home residents in New Jersey during 2001; Tan CG et al.; OBJECTIVE: To characterize risk factors for invasive pneumococcal infection in a nursing home outbreak . DESIGN: Outbreak investigation, case-control study . SETTING: A 114-bed nursing home in New Jersey . PARTICIPANTS: Case-patients were nursing home residents hospitalized with febrile respiratory illness and radiographic findings consistent with pneumonia, and either sputum specimens positive for diplococci or blood cultures positive for Streptococcus pneumoniae, with illness onset during April 3-24, 2001 . Control-patients were selected randomly from remaining residents without respiratory symptoms . METHODS: Chart reviews were performed for case-patients and control-patients . Serotyping and susceptibility testing were performed on S . pneumoniae isolates . Long-term-care facilities (LTCFs) were surveyed to assess compliance with a state regulation mandating pneumococcal vaccination of residents 65 years and older . RESULTS: Nine case-patients were identified, with a median age of 86 years (range, 78 to 100 years) . The median age of control-patients was 86 years (range, 58 to 95 years) . No case-patients versus 9 (50%) control-patients received pneumococcal vaccine before the outbreak (OR, 0; CI95, 0-0.7) . Recent antibiotic use, pneumonia history, and physical functioning were not associated with illness . Illness attack rate was 16% among all unvaccinated residents versus 0 among vaccinated residents . S . pneumoniae serotype 14, included in pneumococcal vaccine, was isolated from blood cultures of 7 case-patients . Of 361 LTCFs (42%) that replied to the survey, 28 (8%) were not complying with state immunization regulations . CONCLUSIONS: This outbreak occurred in an LTCF with low vaccine coverage . Implementing standing order programs, enforcing regulations, documenting vaccinations, and providing education might increase coverage among nursing home residents.

J Dairy Res, 2003 Nov, 70(4), 387 - 94
Effect of penethamate hydriodide treatment on bacteriological cure, somatic cell count and milk production of cows and quarters with chronic subclinical Streptococcus uberis or Streptococcus dysgalactiae infection; St Rose SG et al.; A randomized, controlled field trial was performed in The Netherlands to determine the therapeutic efficacy of parenteral penethamate hydriodide (Leocillin) against naturally occurring, chronic, streptococcal mastitis during lactation . Quarter milk samples were collected from subclinical cases of Streptococcus uberis or Streptococcus dysgalactiae mastitis to determine the effect of treatment on bacteriological cure and somatic cell count (SCC) at quarter level . A quarter was considered to be cured when the bacterial species, isolated prior to treatment, was not isolated from the quarter milk samples taken on days 10 and 20 post-treatment (bacteriological cure), or when a quarter milk SCC (QMSCC) was <250000 cells/ml on days 10 and 20 post-treatment (SCC cure) . Longitudinal data analysis was performed to determine the effect of antibiotic therapy on SCC and milk yield at cow level . Bacteriological cure occurred in 59% of 29 treated quarters, while no cure was observed in any of the 21 untreated control quarters . Treatment resulted in a significant decrease in SCC at cow and quarter level in comparison with untreated controls . There was no significant effect of treatment on milk production . Antibacterial treatment of subclinical streptococcal infections during lactation also prevented clinical mastitis . Furthermore, the treatment may contribute to reduction of bulk milk SCC and to prevention of pathogen spread in dairy herds.

West Indian Med J, 2003 Sep, 52(3), 235 - 9
Infective endocarditis at the University Hospital of the West Indies . A revisit; Coard KC et al.; Many countries have reported a change in the profile of infective endocarditis (IE) over the past three decades . The objective of this study was to evaluate the characteristics of IE from the autopsy service of the University Hospital of the West Indies (UHWI) during the last 15 years and to compare the results with that of an earlier study . There were 26 cases of IE during the period under review with a M:F ratio of 1.2:1 . The ages ranged from 15 days to 74 years with a mean of 36.4 +/- 24.57 years . The largest number of cases (n = 7) occurred in the 10 to 19-year-age group . Cardiac predisposing factors were identified in 14 patients; nine with rheumatic heart disease, four with prosthetic valves and one with a bicuspid aortic valve . All vegetations were located on valves, the aortic being the most frequently involved followed by the mitral . Streptococcus species were the most common causative organisms followed by Staphylococcus aureus . Compared with the profile seen 15 years ago, there have been only minor changes in the characteristics of IE cases observed in the autopsy service at the UHWI.

Vet Ophthalmol, 2003 Dec, 6(4), 309 - 13
Changes in antibiotic resistance in equine bacterial ulcerative keratitis (1991-2000): 65 horses; Sauer P et al.; OBJECTIVE: To document changes in antibiotic resistance of organisms in cases of equine bacterial ulcerative keratitis over a 10-year time period . DESIGN: A retrospective study . PARTICIPANTS: Medical records of equine patients with bacterial ulcerative keratitis seen at the University of Florida's VMTH for the years 1991-2000 were reviewed . MATERIALS AND METHODS: All cases of equine bacterial ulcerative keratitis for the above mentioned years were examined . Bacterial isolates were identified and subjected to Kirby-Bauer disc diffusion method sensitivity tests . Antibiotics used in the sensitivity tests included bacitracin, ampicillin, gentamicin, chloramphenicol, polymyxin B, trimethoprim-sulfa, neomycin, kanamycin, carbenicillin, tobramycin and enrofloxacin . RESULTS: A total of 65 bacterial isolates were subjected to sensitivity testing . Of these isolates, Pseudomonas aeruginosa accounted for 14 of the bacterial isolates (22%), Streptococcus equi subspecies zooepidemicus accounted for 13 of the bacterial isolates (20%), and Staphylococcus aureus accounted for four of the isolates (6%) . A statistically significant increase in resistance of Pseudomonas aeruginosa isolates to the antibiotics gentamicin and tobramycin was found between the isolates from 1992 to 1998 and those from 1999 to 2000 . An increase in resistance of Streptococcus equi subspecies zooepidemicus to gentamicin was found between the isolates from 1993 to 1997 and those from 1998 to 2000 . CONCLUSIONS: Streptococcus equi subspecies zooepidemicus and Pseudomonas aeruginosa were the most common organisms isolated from cases of equine bacterial keratitis referred to the University of Florida's VMTH for the years 1991-2000 . There appears to be an increase in resistance of Streptococcus equi subspecies zooepidemicus to gentamicin over the past 10 years . In addition, there is a significant increase in resistance of Pseudomonas aeruginosa to both gentamicin and tobramycin over the same time period.

J Infect Dis, 2003 Dec 1, 188(11), 1752 - 6 Epub 2003 Nov 07.
Cytokine mRNA expression in pneumococcal carriage, pneumonia, and sepsis in young mice; Ling E et al.; We studied cytokine mRNA expression in Streptococcus pneumoniae carriage, pneumonia, and sepsis in 3-week-old, inbred C57BL/6 and BALB/c mice . Mice were inoculated intranasally with S . pneumoniae serotype 6B, 14, or 3 . Survival, bacterial load in the nasopharynx (NP) and lungs, and mRNA levels of tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, interleukin (IL)-10, and IL-12 in the spleen were analyzed . No baseline mRNA expression levels were found, except for TNF-alpha in C57BL/6 mice . Serotype 6B caused NP colonization only, a significant (P<.05) increase in the levels of TNF-alpha, and induction of TGF-beta and IL-12 mRNA . Serotype 14 caused NP and nonlethal lung colonization and induction of TGF-beta, IL-10, and IL-12 . Serotype 3 caused NP colonization, pneumonia, 35% mortality, and no alteration in the mRNA expression of tested cytokines . Although activation of the immune system culminated in nonlethal disease, evasion of the immune system was associated with detrimental disease.

J Infect Dis, 2003 Dec 1, 188(11), 1679 - 84 Epub 2003 Nov 10.
Epidemiology of acute otitis media caused by Streptococcus pneumoniae before and after licensure of the 7-valent pneumococcal protein conjugate vaccine; McEllistrem MC et al.; We studied, by pulsed-field gel electrophoresis, multilocus sequence typing and penicillin-binding protein 2b amplicon-restriction profiles, pneumococcal isolates recovered from children with acute otitis media during 1 January-31 December 1999 and 2001 . The proportion of nonvaccine serogroups increased from 14.8% (13/88) to 36.5% (23/63) from 1999 to 2001 (P<.01) . Among children who received at least 2 doses of the pneumococcal 7-valent protein conjugate (PNC7) vaccine, 46.7% (7/15) of the isolates had nonvaccine serogroups, compared with 20.8% (26/125) of the isolates from children who did not receive the PNC7 vaccine (P=.05) . Overall, the serogroups involved in capsular switching were 6-19-NT, 6-14-35, 15-19, and the 19-Spanish 23F clone . In 1999 and 2001, 30.8% (4/13) and 26.1% (6/23) of the nonvaccine serogroups were implicated in capsular switching, respectively . Continued surveillance will be of great importance as the distribution of the PNC7 vaccine increases.

Am J Infect Control, 2003 Nov, 31(7), 410 - 4
Investigation of antimicrobial use pattern in the intensive treatment unit of a teaching hospital in western Nepal; Shankar PR et al.; BACKGROUND: Inappropriate use of antimicrobials is of special importance in the intensive treatment unit because of the large number of drugs prescribed, the chance for drug errors, and the likelihood of development of drug resistance . METHODS: A total of 297 records of patients admitted to the intensive treatment unit of the Manipal teaching hospital, a tertiary care hospital in Pokhara, western Nepal, were studied to determine the prescribing frequency and rationality of use of antimicrobials . Patient outcome, duration of stay in the intensive treatment unit, and the age and sex distribution of the patients were also studied . RESULTS: Mean+/-SD drugs per patient was 3.4+/-1.8 . About half (50.2%) of the patients received an antimicrobial; 84.6% of the antimicrobials were used without obtaining bacteriologic evidence of infection . The commonest organisms isolated on culture were Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, and Staphylococcus aureus . A total of 28.9% of the antimicrobials were prescribed for lower respiratory tract infections on the basis of the putative site of infection; 61.9% of the antimicrobials were prescribed by the parenteral route and mainly the older generation of antimicrobials were used . In 39 of the 149 patients prescribed an antimicrobial, the use was irrational . CONCLUSIONS: Prescriber education to improve prescribing patterns and regular auditing of antimicrobial prescriptions to prevent their inappropriate use and unnecessary cost to the patients are required . The high percentage of inappropriate use of antimicrobials raises concerns about the development and spread of drug resistance, which must be addressed.

Infect Immun, 2003 Dec, 71(12), 7197 - 201
Identification of B- and T-cell epitopes within the fibronectin-binding domain of the SfbI protein of Streptococcus pyogenes; Schulze K et al.; The fibronectin-binding repeats of the SfbI protein of Streptococcus pyogenes constitute the minimal domain able to confer protection against lethal infection . We investigated the presence of B- and T-cell epitopes within this region in congenic mice . One linear B-cell epitope was recognized by BALB/b and BALB/k mice, whereas two epitopes were found in BALB/c animals . A unique T-cell epitope was recognized by all three mouse strains . All identified epitopes clustered in a 30-amino-acid fragment . These results suggest that this polypeptide may be suitable for incorporation into a polyepitope-based vaccine formulation against S . pyogenes.

Infect Immun, 2003 Dec, 71(12), 7193 - 6
The activation of bovine plasminogen by PauA is not required for virulence of Streptococcus uberis; Ward PN et al.; A mutant of Streptococcus uberis carrying a single copy of ISS1 within pauA was unable to activate bovine plasminogen . Contrary to a hypothesis postulated previously, this mutation did not alter the ability of the bacterium to grow in milk or to infect the lactating bovine mammary gland.

Infect Immun, 2003 Dec, 71(12), 7149 - 53
Pneumococcal surface protein A is expressed in vivo, and antibodies to PspA are effective for therapy in a murine model of pneumococcal sepsis; Swiatlo E et al.; Pneumococcal surface protein A (PspA) is an immunogenic protein expressed on the surface of all strains of Streptococcus pneumoniae (pneumococcus) and induces antibodies which protect against invasive infection in mice . Pneumococci used for infectious challenge in protection studies are typically collected from cultures grown in semisynthetic medium in vitro . The purpose of these studies is to confirm that PspA is expressed by pneumococci during growth in vivo at a level sufficient for antibodies to PspA to be protective . Mice were actively immunized with purified PspA or by passive transfer of monoclonal antibody (MAb) and challenged with a capsular type 3 strain in diluted whole blood from bacteremic mice . All were protected against challenge with 10 times the 50% lethal dose (LD(50)), and mice challenged with 1,000 times the LD(50) had increased survival compared with controls . Additionally, nonimmune mice treated with MAbs to PspA or PspA immune serum at 6 and 12 h after infection with 10 times the LD(50) also showed increased survival . Northern blot analysis of RNA from pneumococci grown either in vitro or in vivo showed similar levels of PspA mRNA . These results demonstrate that PspA is expressed in vivo in a mouse model and that immunization with PspA induces antibodies to an antigen which is expressed during the course of invasive infection . Immunotherapy with antibodies to PspA may have some utility in treating pneumococcal infections in humans.

Infect Immun, 2003 Dec, 71(12), 7079 - 86
Prophage induction and expression of prophage-encoded virulence factors in group A Streptococcus serotype M3 strain MGAS315; Banks DJ et al.; The genome of the highly virulent group A Streptococcus (GAS) serotype M3 strain MGAS315 has six prophages that encode six proven or putative virulence factors . We examined prophage induction and expression of prophage-encoded virulence factors by this strain under in vitro conditions inferred to approximate in vivo conditions . Coculture of strain MGAS315 with Detroit 562 (D562) human epithelial pharyngeal cells induced the prophage encoding streptococcal pyrogenic exotoxin K (SpeK) and extracellular phospholipase A(2) (Sla) and the prophage encoding streptodornase (Sdn) . Increased gene copy numbers after induction correlated with increased speK, sla, and sdn transcript levels . Although speK and sla are located contiguously in prophage Phi315.4, these genes were transcribed independently . Whereas production of immunoreactive SpeK was either absent or minimal during coculture of GAS with D562 cells, production of immunoreactive Sla increased substantially . In contrast, despite a lack of induction of the prophage encoding speA during coculture of GAS with D562 cells, the speA transcript level and production of immunoreactive streptococcal pyrogenic exotoxin A (SpeA) increased . Exposure of strain MGAS315 to hydrogen peroxide, an oxidative stressor, induced the prophage encoding mitogenic factor 4 (MF4), and there was a concomitant increase in the mf4 transcript . All prophages of strain MGAS315 that encode virulence factors were induced during culture with mitomycin C, a DNA-damaging agent . However, the virulence factor gene transcript levels and production of the encoded proteins decreased after mitomycin C treatment . Taken together, the results indicate that a complex relationship exists among environmental culture conditions, prophage induction, and production of prophage-encoded virulence factors.

Infect Immun, 2003 Dec, 71(12), 6707 - 11
Oxygen regulates invasiveness and virulence of group B streptococcus; Johri AK et al.; The facultative anaerobe group B Streptococcus (GBS) is an opportunistic pathogen of pregnant women, newborns, and the elderly . Although several virulence factors have been identified, environmental factors that regulate the pathogenicity of GBS have not been well characterized . Using the dynamic in vitro attachment and invasion system (DIVAS), we examined the effect of oxygen on the ability of GBS to invade immortalized human epithelial cells . GBS type III strain M781 invaded human epithelial cells of primitive neurons, the cervix, the vagina, and the endometrium in 5- to 400-fold higher numbers when cultured at a cell mass doubling time (t(d)) of 1.8 h than at a slower t(d) of 11 h . Invasion was optimal when GBS was cultured at a t(d) of 1.8 h in the presence of >or=5% oxygen and was significantly reduced without oxygen . Moreover, GBS grown in a chemostat under highly invasive conditions (t(d) of 1.8 h, with oxygen) was more virulent in neonatal mice than was GBS grown under suboptimal invasion conditions (t(d) of 1.8 h, without oxygen), suggesting a positive association between in vitro invasiveness with DIVAS and virulence.

Antimicrob Agents Chemother, 2003 Dec, 47(12), 3935 - 41
Pharmacodynamics of the new des-f(6)-quinolone garenoxacin in a murine thigh infection model; Andes D et al.; Garenoxacin is a new des-F(6)-quinolone with broad-spectrum activity against both gram-positive cocci and gram-negative bacilli . We used the neutropenic murine thigh infection model to characterize the time course of antimicrobial activity of garenoxacin and determine which pharmacokinetic-pharmacodynamic (PK-PD) parameter best correlated with efficacy . Serum drug levels following three fourfold-escalating single-dose levels of garenoxacin were measured by microbiologic assay . In vivo postantibiotic effects (PAEs) were determined after doses of 16 and 64 mg/kg of body weight . Mice had 10(6.5) to 10(6.7) CFU of Streptococcus pneumoniae strain ATCC 10813 or Staphylococcus aureus strain ATCC 33591 per thigh when they were treated for 24 h with garenoxacin at a dose of 4 to 128 mg/kg/day fractionated for 3-, 6-, 12-, and 24-hour dosing regimens . Nonlinear regression analysis was used to determine which PK-PD parameter best correlated with the measurement of CFU/thigh at 24 h . Pharmacokinetic studies yielded peak/dose values of 0.2 to 0.3, area under the concentration-time curve (AUC)/dose values of 0.1 to 0.5, and half-lives of 0.7 to 1.6 h . Garenoxacin produced in vivo PAEs of 1.4 to 8.2 h with S . pneumoniae ATCC 10813, 7.6 to >12.4 h with S . aureus ATCC 25923, and 0 to 1.5 h with Klebsiella pneumoniae ATCC 43816 . The 24-h AUC/MIC ratio was the PK-PD parameter that best correlated with efficacy (R2=71 to 90% for the two organisms compared with 43 to 56% for the peak/MIC ratio and 47 to 75% for percent time above the MIC {% T>MIC}) . In subsequent studies we used the neutropenic murine thigh infection model to determine if the magnitude of the AUC/MIC ratio needed for efficacy of garenoxacin varied among pathogens (including resistant strains) . Mice had 10(5.9) to 10(7.2) CFU of 6 strains of S . aureus (2 methicillin resistant), 11 strains of S . pneumoniae (5 penicillin susceptible, 1 penicillin intermediate, and 5 penicillin resistant, and of the resistant strains, 3 were also ciprofloxacin resistant), and 4 gram-negative strains per thigh when treated for 24 h with 1 to 64 mg of garenoxacin per kg every 12 h . A sigmoid dose-response model was used to estimate the doses (mg/kg/24 h) required to achieve a net bacteriostatic effect over 24 h . MICs ranged from 0.008 to 4 microg/ml . The free drug 24-h AUC/MIC ratios for each static dose (2.8 to 128 mg/kg/day) varied from 8.2 to 145 . The mean 24-h AUC/MIC ratios +/- standard deviations for S . pneumoniae, S . aureus, and gram-negative strains were 33 +/- 18, 81 +/- 37, and 33 +/- 30, respectively . Methicillin, penicillin, or ciprofloxacin resistance did not alter the magnitude of the AUC/MIC ratio required for efficacy.

Antimicrob Agents Chemother, 2003 Dec, 47(12), 3699 - 703
In vivo efficacy of a new quinolone, DQ-113, against Streptococcus pneumoniae in a mouse model; Otsu Y et al.; DQ-113 is a new quinolone with potent activity against gram-positive pathogens . The in vivo activity of DQ-113 against Streptococcus pneumoniae was compared with those of gatifloxacin and ciprofloxacin in a mouse model . For this purpose, two strains of S . pneumoniae were used: penicillin-susceptible S . pneumoniae (PSSP) and penicillin-resistant S . pneumoniae (PRSP) . The survival rates of mice infected with PSSP and PRSP at 14 days after infection were 80% in the DQ-113-treated group and 0 to 10% in the other three groups . In murine infections caused by PSSP, the 50% effective doses (ED50s) of DQ-113, gatifloxacin, and ciprofloxacin were 6.0, 41.3, and 131.6 mg/kg, respectively . Against PRSP-caused pneumonia in mice, the ED50s of DQ-113, gatifloxacin, and ciprofloxacin were 7.6, 64.7, and 125.9 mg/kg, respectively . Compared with the other drugs, DQ-113 showed excellent therapeutic efficacy and eradicated viable bacteria in both PSSP- and PRSP-infected mice . The means +/- standard errors of the means of viable bacterium counts in the lungs of gatifloxacin-treated, ciprofloxacin-treated, and untreated control mice infected with PSSP were 2.91 +/- 0.34, 3.13 +/- 0.48, and 3.86 +/- 0.80 log10 CFU/ml, respectively . The same counts in mice infected with PRSP treated with the same three agents were 6.57 +/- 0.99, 6.54 +/- 0.40, and 7.17 +/- 0.43 log10 CFU/ml, respectively . DQ-113 significantly decreased the number of viable bacteria in the lungs compared with gatifloxacin and ciprofloxacin . Of the drugs analyzed, the pharmacokinetic-pharmacodynamic parameter of area under the concentration-time curve (AUC)/MIC ratio for DQ-113 was significantly higher than those for gatifloxacin and ciprofloxacin . Our results suggest that DQ-113 has potent in vivo efficacy against both PSSP and PRSP.

Vet Microbiol, 2003 Dec 2, 97(1-2), 135 - 51
Epidemiology of Streptococcus suis serotype 5 infection in a pig herd with and without clinical disease; Cloutier G et al.; The aim of this study was to describe the transmission and the kinetics of the infection caused by Streptococcus suis serotype 5 in a multisite farrow-to-finish pig herd . Most sows carried S . suis serotype 5 in their vaginal tract, but not in their nasal cavities, as demonstrated by immunomagnetic separation (IMS) technique . Their offspring became infected during farrowing, confirming vertical transmission . During the first 4 weeks of life, a low number of piglets were carriers of S . suis serotype 5 in their nasal cavities . However, when clinical signs appeared, the carrier rate significantly increased, suggesting that isolation from nasal cavities is a better indication of active transmission than of a carrier state . Clinical cases were present in animals between 4 and 8 weeks of age, when maternal antibodies were at their lowest level . Up to six different genotypes of the same serotype could be identified by random amplified polymorphic DNA; however, a single clone was responsible for all clinical cases studied . This clone could only be isolated from a single sow, indicating that its prevalence in breeding animals was low . Interestingly, 1 year later, clinical disease associated with S . suis serotype 5 spontaneously disappeared . At that time, the genotype responsible for the clinical signs was not detected in the herd and the levels of antibodies in sows and maternal antibodies in piglets were not higher than those of the previous year.

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