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 ea