J Infect Dis, 1999 Sep, 180(3), 888 - 91 Fcgamma receptor polymorphisms determine the magnitude of in vitro phagocytosis of Streptococcus pneumoniae mediated by pneumococcal conjugate sera; Jansen WT et al.; Fcgamma receptors show two genetically determined polymorphisms: the biallelic FcgammaRIIa-R131 and -H131 polymorphism and the NA1/NA2 FcgammaIIIb polymorphism . Using 10 pre- and postconjugate vaccination sera from adults, we analyzed in vitro phagocytic capacities of three different combinations of polymorphonuclear leukocyte FcgammaR allotypes: those homozygous for the H131 and NA1 allotype, those homozygous for the R131 and NA2 allotype, and those heterozygous for both receptors . For pre- and postvaccination sera, mean phagocytosis levels for the homozygous H131/NA1 allotype were 4 -fold higher than for the homozygous R131/NA2 allotype . There was a strong and significant correlation between IgG2 ELISA antibody titers and phagocytosis levels for the homozygous H131/NA1 Fcgamma receptor allotype and the heterozygous allotype but not for the homozygous R131/NA2 allotype . There was no relation between IgG1 ELISA titer and phagocytosis level . Apparently the IgG2 antibodies induced are functionally the most important . This may explain the large effect of Fcgamma receptor polymorphisms on in vitro phagocytosis of pneumococci mediated by conjugate antisera. Wkly Epidemiol Rec, 1999 Jun 11, 74(23), 177 - 83 Pneumococcal vaccines . WHO position paper; Fatal group A Streptococcal toxic shock-like syndrome in a child with varicella: report of the first well documented case with detection of the genetic sequences that code for exotoxins spe A and B et al.; Intensive Care Unit, Instituto de Infectologia Emilio Ribas, Sao Paulo, Brazil . jaquess@icr.hcnet.usp.br A previously healthy seven-year-old boy was admitted to the intensive care unit because of toxaemia associated with varicella . He rapidly developed shock and multisystem organ failure associated with the appearance of a deep-seated soft tissue infection and, despite aggressive treatment, died on hospital day 4 . An M-non-typable, spe A and spe B positive Group A Streptococcus was cultured from a deep soft tissue aspirate . The criteria for defining Streptococcal toxic shock-like syndrome were fulfilled . The authors discuss the clinical and pathophysiological aspects of this disease as well as some unusual clinical findings related to this case. Rev Hosp Clin Fac Med Sao Paulo, 1998 May-Jun, 53(3), 152 - 5 {Early-onset neonatal sepsis and late-appearing diaphragmatic hernia}; Falcao MC et al.; The interesting association between delayed presentation of right-sided diaphragmatic hernia and neonatal Group B streptococcal infections occurs rarely and its pathogenesis is still obscure . Two preterm newborn infants with early onset of neonatal sepsis (one due to Group B Streptococcus) followed by recognition of right-sided diaphragmatic hernia on the 9th and 25th day of life are reported . In both cases the course of neonatal sepsis and pneumonia was complicated due to the appearance of right-sided pleural effusion and atelectasis . On serial chest roentgenograms right-sided bowel gas was noticed and the liver shadow became gradually elevated . Diagnosis was confirmed by ultrasonography and computed tomography . Suspicion of associated diaphragmatic hernia should be raised in neonatal streptococcal infection whenever subsequent progressive respiratory deterioration ensues, requiring mechanical ventilation after initial clinical improvement or in the presence of right-sided pleural effusion. Ann Otol Rhinol Laryngol, 1999 Jul, 108(7 Pt 1), 648 - 52 Expression of intercellular adhesion molecule-1 in rat inner ear due to bacterial otitis media; Sone M et al.; Expressions of anti-Streptococcus pneumoniae antibody and anti-intercellular adhesion molecule-1 (ICAM-1) antibody in the inner ear were studied immunohistochemically in rats following inoculation of S pneumoniae type 6A into the middle ear cavity . Positive staining with anti-S pneumoniae antibody was detected in the marginal cells of the stria vascularis of rats sacrificed 3 days after S pneumoniae inoculation, but almost no staining was detected in those sacrificed at 14 days . Strong ICAM-1 expression was detected in the basal cell layer of the stria vascularis of rats sacrificed 3 days after inoculation, but the intensity of the staining had decreased by 14 days . These results suggest that the stria vascularis may be a site of the inner ear damage that follows bacterial inoculation into the middle ear cavity . The up-regulated expression of ICAM-1 in the basal cell layer may represent a reaction of the inner ear to the bacterial otitis media. Ann Otol Rhinol Laryngol, 1999 Jul, 108(7 Pt 1), 629 - 33 Documentation of the prevalence of penicillin-resistant Streptococcus pneumoniae isolated from the middle ear and sinus fluid of children undergoing tympanocentesis or sinus lavage; Shapiro NL et al.; With increasing pneumococcal resistance to penicillin and other antibiotics, use of antibiotic therapy for children with upper respiratory tract infections such as otitis media and sinusitis has become difficult . Selecting an appropriate treatment regimen has become more challenging due to frequent concomitant microbial resistance to multiple antibiotics . In a prospective, nonrandomized study, we obtained middle ear and sinus aspirate specimens from all children undergoing outpatient tympanocentesis or sinus lavage for any indication at our institution over two 4-week periods . One hundred fifty-four specimens were obtained . Of these, 12 grew Streptococcus pneumoniae, 7 of which were resistant to penicillin . A 6-month retrospective review of these patients' medical histories evaluated their antibiotic use prior to surgical intervention . An association between penicillin resistance and recent use of 2 or more antibiotics in children with positive S pneumoniae cultures was confirmed, as has been documented in prior reports . Those with penicillin-resistant S pneumoniae also demonstrated a higher incidence of multidrug-resistant organisms. Bone Marrow Transplant, 1999 Jul, 24(1), 89 - 93 Totally implantable central venous access ports for high-dose chemotherapy administration and autologous stem cell transplantation: analysis of overall and septic complications in 68 cases using a single type of device; Biffi R et al.; Sixty-eight patients suffering from breast cancer, ovarian cancer, lymphoma or multiple myeloma were treated with high-dose chemotherapy and autologous stem cell transplantation . They underwent placement of a central venous port via the subclavian vein for delivery of chemotherapy and reinfusion of stem cells . All patients were followed prospectively for device-related and overall complications, comprising a total of 18,213 days in situ (median: 267 days, range: 90-480) . One patient experienced a pneumothorax (1.4%) spontaneously resolved, as an acute toxicity . Two patients (2.8%, 0.1 episodes/1000 days of use) were forced to have the port removed due to infection, caused by Streptococcus mitis in one case, while the causative agent was not identified by laboratory tests in the second . The other 66 patients completed the therapeutic programme, including peripheral stem cell reinfusions and supportive care, such as i.v . antibiotics, antiemetics or fluid administration and blood sample collection, without additional complications . In conclusion, the use of totally implantable central venous access ports has resulted in good long-term access to central veins, in spite of the severe neutropenia and increased septic risk of this category of oncology patients. J Chemother, 1999 Jun, 11(3), 191 - 4 Susceptibility of penicillin-resistant and penicillin-susceptible Streptococcus pneumoniae to newer antimicrobial agents; Rubio MC et al.; Agar dilution minimum inhibitory concentration (MIC) methodology, according to NCCLS guidelines, was used to test the activity of three glycopeptides (LY 333328 {LY}, vancomycin {VAN}, and teicoplanin {TEI}), four fluoroquinolones (trovafloxacin {TRO}, BAY 12-8039 {BAY}, ciprofloxacin {CIP}, and ofloxacin {OFL}), five macrolide-lincosamide-streptogramin antibiotics (erythromycin {ERY}, azithromycin {AZI}, miocamycin {MOM}, clindamycin CLN}, and quinupristin-dalfopristin {SYN} against 126 Streptococcus pneumoniae strains, isolated in Lozano Blesa Hospital of Zaragoza (Spain) . MIC50/MIC90 (microg/ml) values for penicillin-susceptible (PS), penicillin-intermediate (PI) and penicillin-resistant (PR) strains show an excellent antipneumococcal activity of LY 333326--a new glycopeptide, for the fluoroquinolones trovafloxacin and moxifloxacin {BAY 12-8039}, and for quinupristin/dalfopristin, regardless of the resistance phenotype of the strains. J Fr Ophtalmol, 1999 Jun-Jul, 22(6), 662 - 5 {Use of topical cyclosporin in microcrystalline keratopathy due to Streptococcus}; Touzeau O et al.; Infectious crystalline keratopathy is an exceptional but serious complication of penetrating keratoplasty . Streptococcus viridans and a steroid treatment are very often found . The treatment is always lengthy and difficult . The high number of bacteria and the steroid treatment, necessary to prevent rejection, worsen the infection . We report a case of Streptococcus crystalline keratopathy that evolved, despite the treatment, simultaneously toward abscess and acute rejection . Topical cyclosporin treatment was used to maintain an immunosuppression without worsening the infection. Clin Infect Dis, 1999 Jul, 29(1), 196 - 8 Endocarditis caused by group A beta-hemolytic Streptococcus in an infant: case report and review; Winterbotham A et al.; Acute bacterial endocarditis in the absence of underlying heart disease is rare . We report the occurrence of endocarditis caused by group A beta-hemolytic Streptococcus (GABHS), following varicella, in a 5-month-old child without heart disease . In addition to this child, seven other children with endocarditis caused by GABHS have been reported since 1966, six of whom did not have preexisting heart disease . In one of these children, GABHS endocarditis was preceded by varicella . These cases indicate that GABHS is capable of causing endocarditis in the absence of heart disease, and they provide further evidence that varicella is an important risk factor for invasive GABHS infections. J Ethnopharmacol, 1999 Aug, 66(2), 235 - 40 Preliminary screening of ethnomedicinal plants from India; Perumal Samy R et al.; Antibacterial activity of aqueous residues of 16 different ethnomedicinal plants have been studied . The effect of the aqueous extract at two different weights of plant residues, 30 and 40 mg, were tested against three gram positive bacteria and seven gram negative bacteria by the filter paper disc diffusion method . Among the tested plants, Cleome gynandropsis and Ageratum conyzoides showed a significant control of the growth of Alkaligens viscolactis, Klebsiella aerogenas, Bacillus cerues and Streptococcus pyogens . The maximum inhibitions were observed in Tridax procumbens, Cleome viscosa, Acalypha indica and Boerhaavia erecta against Aeromonas hydrophilla and Bacillus cerues. J Exp Med, 1999 Jul 19, 190(2), 241 - 51 A single gene (tts) located outside the cap locus directs the formation of Streptococcus pneumoniae type 37 capsular polysaccharide . Type 37 pneumococci are natural, genetically binary strains; Llull D et al.; The molecular aspects of the type 37 pneumococcal capsular biosynthesis, a homopolysaccharide composed of sophorosyl units (beta-d-Glc-(1-->2)-beta-d-Glc) linked by beta-1,3 bonds, have been studied . Remarkably, the biosynthesis of the type 37 capsule is driven by a single gene (tts) located far apart from the cap locus responsible for capsular formation in all of the types characterized to date in Streptococcus pneumoniae . However, a cap37 locus virtually identical to the cap33f cluster has been found in type 37 strains, although some of its genes are inactivated by mutations . The tts gene has been sequenced and its transcription start point determined . Tts shows sequence motifs characteristic of cellulose synthases and other beta-glycosyltransferases . Insertion of the tts gene into the pneumococcal DNA causes a noticeable genome reorganization in such a way that genes normally separated by more than 350 kb in the chromosome are located together in clinical isolates of type 37 . Encapsulated pneumococcal strains belonging to 10 different serotypes (or serogroups) transformed with tts synthesized type 37 polysaccharide, leading to the formation of strains that display the binary type of capsule . Type 37 pneumococcus constitutes the first case of a natural, genetically binary strain and represents a novel alternative to the mechanisms of intertype transformation. Am J Respir Crit Care Med, 1999 Aug, 160(2), 672 - 7 Bacterial colonization of central airways after stenting; Noppen M et al.; Airway stenting (AS) is increasingly used in the management of obstructive lesions of the central airways . Although retention of secretions and infection have been reported as complications of AS, the microbiological consequences of AS have not yet been evaluated . In this study, we prospectively performed protected specimen brush (PSB) sampling of the airways, before and 3 to 4 wk after AS, in 14 consecutive patients (65 +/- 17 yr), suffering from bronchial (5), extensive esophageal (2), thyroid (1), and adenocystic (1) carcinoma, stenotic tracheal burn lesions (2), postintubation stenosis (2), and Wegener's granulomatosis (1) . A cutoff value of >/= 10(2) colony-forming units (cfu) . ml(-)(1) was considered diagnostic for airway colonization (AC) . PSB results were related to the presence and degree of secretion retention (SR) at the level of the stent . In five of the 14 patients, AC was present prior to AS; in three of these, potentially pathogenic microorganisms (PPM) were identified . After AS, AC was found in 11 (including seven patients without prior AC) of the 14 patients . In six of these patients, one or more PPM were present (Pseudomonas aeruginosa {4}, Staphylococcus aureus {3}, Streptococcus pneumoniae {1}, Klebsiella spp . {1}) . Although AC tended to be associated with the presence of SR (PSB >/= 10(2) cfu . ml(-)(1) in 10 of 12 SR-positive and in zero SR-negative cases; PSB < 10(2) cfu . ml(-)(1) in two SR-positive and in two SR-negative cases), statistical significance was not reached (Fisher exact test, p = 0.06) . We conclude that AS is frequently followed by AC, the majority of which occurs in patients without AC prior to AS, and is caused by PPM . In no case, however, AC was associated with clinical signs of infection . AC tended to be associated with SR in the stent. Hepatogastroenterology, 1999 May-Jun, 46(27), 1782 - 4 Streptococcus milleri infection and pericardial abscess associated with esophageal carcinoma: report of two cases; Muto M et al.; We report 2 cases of esophageal carcinoma with esophago-mediastinal fistula that developed pericardial abscess due to Streptococcus intermedius infection . Streptococcus intermedius, a generally harmless commensals in healthy humans, is not usually associated with infections of the oral cavity but may account for non-oral purulent infections . This report, however, highlights that Streptococcus intermedius infection can be life-threatening for some patients such as those with esophageal carcinoma with fistula. Eur J Biochem, 1999 Jul, 263(1), 145 - 51 Autocatalytic processing of the streptococcal cysteine protease zymogen: processing mechanism and characterization of the autoproteolytic cleavage sites; Doran JD et al.; The autocatalytic processing of the streptococcal cysteine protease zymogen (proSCP) to active streptococcal cysteine protease (SCP) was investigated in vitro using purified protein from Streptococcus pyogenes strain B220 . It was found that the autocatalytic maturation of the zymogen proceeds through the sequential appearance of at least six intermediates, five of which were characterized through a combination of N-terminal sequencing and MS . Intermediates were identified as resulting from cleavages after Lys26, Asn41, Lys101, Ala112, and Lys118 . Time-course studies of the proSCP processing gave a sigmoidal activity profile and indicated that proSCP catalyses its own transformation, mainly via an intermolecular processing mechanism . A similar sequential appearance of intermediates was observed when inactive Cys192Ser proSCP was treated with native, enzymatically active SCP, thus demonstrating that the maturation can exclusively proceed by a bimolecular mechanism . It was shown that proSCP, but not mature SCP, immobilized on a Sepharose resin is capable of liberating itself from the column, indicating that the zymogen is also capable of intramolecular processing . In order to test whether the amino acid sequences at the processing sites could be used for developing new, specific substrates, 3-amino benzoic acid octapeptide derivatives based on all five characterized amino acid sequences from the autoprocessing cleavage sites were synthesized and tested for activity . The 3-amino benzoic acid derivatives have kcat/KM values ranging from 1200 to 7700.M-1.s-1, making them very good endopeptidase substrates for SCP. Antimicrob Agents Chemother, 1999 Aug, 43(8), 2005 - 9 Evaluation of low-dose, extended-interval clindamycin regimens against Staphylococcus aureus and Streptococcus pneumoniae using a dynamic in vitro model of infection; Lewis RE et al.; We have previously described the activity of low-dose clindamycin in extended-interval dosing regimens by determination of bactericidal titer in serum . In this study, we used a one-compartment in vitro dynamic infection model to compare the pharmacodynamics of clindamycin in three intravenous-dosing regimens (600 mg every 8 h {q8h}, 300 mg q8h, and 300 mg q12h) against three clinical isolates of Staphylococcus aureus and two clinical isolates of Streptococcus pneumoniae . Test organisms were added to the central compartment of the model to yield a starting inoculum of 10(6) CFU/ml . Clindamycin was injected as a bolus into the central compartment at appropriate times over 48 h to simulate the q8h or q12h dosing regimens . Drug-free culture medium was then pumped through the system to mimic a half-life of 2.4 h . At predetermined time points during the experiment, samples were removed from the central compartments for colony count determination and drug concentration analysis . The rates of killing did not significantly differ among the three clindamycin dosing regimens against either S . aureus or S . pneumoniae (P = 0.88 or 0.998, respectively) . Likewise, no significant differences in activities were detected among the three regimens against staphylococci (P = 0.677 and 0.667) or pneumococci (P = 0.88 and 0.99) . Against an S . aureus isolate exhibiting inducible macrolide-lincosamide-streptogramin B resistance, none of the three clindamycin regimens prevented regrowth of the resistance phenotype in the model . In this model, clindamycin administered at a low dose in an extended-interval regimen (300 mg q12h) exhibited antibacterial activity equivalent to that of the 300- or 600-mg-q8h regimen. Antimicrob Agents Chemother, 1999 Aug, 43(8), 2000 - 4 Contribution of topoisomerase IV and DNA gyrase mutations in Streptococcus pneumoniae to resistance to novel fluoroquinolones; Pestova E et al.; In this study, we assessed the activity of ciprofloxacin, levofloxacin, sparfloxacin, and trovafloxacin against clinical isolates of Streptococcus pneumoniae that were resistant to the less-recently developed fluoroquinolones by using defined amino acid substitutions in DNA gyrase and topoisomerase IV . The molecular basis for resistance was assessed by using mutants selected with trovafloxacin, ciprofloxacin, and levofloxacin in vitro . This demonstrated that the primary target of trovafloxacin in S . pneumoniae is the ParC subunit of DNA topoisomerase IV, similar to most other fluoroquinolones . However, first-step mutants bearing the Ser79-->Phe/Tyr substitution in topoisomerase IV subunit ParC were susceptible to trovafloxacin with a minimum inhibitory concentration of 0.25 microg/ml, and mutations in the structural genes for both topoisomerase IV subunit ParC (parC) and the DNA gyrase subunit (gyrA) were required to achieve levels of resistance above the breakpoint . The data also suggest that enhanced activity of trovafloxacin against pneumococci is due to a combination of factors that may include reduced efflux of this agent and an enhanced activity against both DNA gyrase and topoisomerase IV. Antimicrob Agents Chemother, 1999 Aug, 43(8), 1935 - 40 Phenotypes and genotypes of erythromycin-resistant Streptococcus pyogenes strains in Italy and heterogeneity of inducibly resistant strains; Giovanetti E et al.; A total of 387 clinical strains of erythromycin-resistant (MIC, >/=1 microg/ml) Streptococcus pyogenes, all isolated in Italian laboratories from 1995 to 1998, were examined . By the erythromycin-clindamycin double-disk test, 203 (52.5%) strains were assigned to the recently described M phenotype, 120 (31.0%) were assigned to the inducible macrolide, lincosamide, and streptogramin B resistance (iMLS) phenotype, and 64 (16.5%) were assigned to the constitutive MLS resistance (cMLS) phenotype . The inducible character of the resistance of the iMLS strains was confirmed by comparing the clindamycin MICs determined under normal testing conditions and those determined after induction by pregrowth in 0.05 microg of erythromycin per ml . The MICs of erythromycin, clarithromycin, azithromycin, josamycin, spiramycin, and the ketolide HMR3004 were then determined and compared . Homogeneous susceptibility patterns were observed for the isolates of the cMLS phenotype (for all but one of the strains, HMR3004 MICs were 0.5 to 8 microg/ml and the MICs of the other drugs were >128 microg/ml) and those of the M phenotype (resistance only to the 14- and 15-membered macrolides was recorded, with MICs of 2 to 32 microg/ml) . Conversely, heterogeneous susceptibility patterns were observed in the isolates of the iMLS phenotype, which were subdivided into three distinct subtypes designated iMLS-A, iMLS-B, and iMLS-C . The iMLS-A strains (n = 84) were highly resistant to the 14-, 15-, and 16-membered macrolides and demonstrated reduced susceptibility to low-level resistance to HMR3004 . The iMLS-B strains (n = 12) were highly resistant to the 14- and 15-membered macrolides, susceptible to the 16-membered macrolides (but highly resistant to josamycin after induction), and susceptible to HMR3004 (but intermediate or resistant after induction) . The iMLS-C strains (n = 24) had lower levels of resistance to the 14- and 15-membered macrolides (with erythromycin MICs increasing two to four times after induction), were susceptible to the 16-membered macrolides (but resistant to josamycin after induction), and remained susceptible to HMR3004, also after induction . The erythromycin resistance genes in 100 isolates of the different groups were investigated by PCR . All cMLS and iMLS-A isolates tested had the ermAM (ermB) gene, whereas all iMLS-B and iMLS-C isolates had the ermTR gene (neither methylase gene was found in isolates of other groups) . The M isolates had only the macrolide efflux (mefA) gene, which was also found in variable proportions of cMLS, iMLS-A, iMLS-B, and iMLS-C isolates . The three iMLS subtypes were easily differentiated by a triple-disk test set up by adding a josamycin disk to the erythromycin and clindamycin disks of the conventional double-disk test . Tetracycline resistance was not detected in any isolate of the iMLS-A subtype, whereas it was observed in over 90% of both iMLS-B and iMLS-C isolates. Eur J Immunol, 1999 Jul, 29(7), 2297 - 308 Design of highly immunogenic liposomal constructs combining structurally independent B cell and T helper cell peptide epitopes; Boeckler C et al.; We have designed liposomal diepitope constructs that allow the physical combination, within the same vesicle, of B and Th epitopes as structurally separate entities . The immune response against such constructs was explored using TPEDPTDPTDPQDPSS (TPE), a B cell epitope originating from a Streptococcus mutans surface adhesin and QYIKANSKFIGITEL (QYI), a "universal" Th epitope from tetanus toxin . The two peptides were linked to the outer surface of small (diameter approximately 100 nm) unilamellar liposomes by covalent conjugation to two different anchors . To that end we have developed a strategy that allows the controlled chemical coupling of TPE and QYI, functionalized at their N terminus with a thiol, to preformed liposomes containing thiol-reactive derivatives of phosphatidylethanolamine and the lipopeptide S-{2,3-bis (palmitoyloxy)-(2-RS)-propyl}-N-palmitoyl-(R)-cysteinyl-alanyl-gly cine (Pam3CAG), respectively . This synthetic construct (administered i.p . to BALB/c mice) induced highly intense (titers > 20,000), anamnestic and long-lasting (over 2 years) immune responses, indicating that this strategy is successful . Two parameters were of prime importance to elicit this response with our liposomal diepitope constructs: (1) the simultaneous expression of B and Th epitopes on the same vesicle, and (2) the lipopeptide Pam3CAG anchor of the Th epitope QYI could not be replaced by a phosphatidylethanolamine anchor (a lesser immune response was observed) . Analysis of the antibody response revealed a complex pattern; thus, besides the humoral response (production of IgG1, IgG2a, IgG2b) a superposition of a T-independent (TI-2 type) response was also found (IgM and IgG3) . These results indicate that liposomal diepitope constructs could be attractive in the development of synthetic peptide-based vaccines. FEMS Microbiol Lett, 1999 Jul 15, 176(2), 421 - 8 Survival of Streptococcus pyogenes under stress and starvation; Trainor VC et al.; The ability of Streptococcus pyogenes to enter a quiescent state, similar to the stationary phase of lab cultures, is believed to be an important factor in its ability to persist within the host and to subsequently cause disease . Using a model broth system, we determined that after entering the stationary phase, there was a 99.99% reduction in cell viability over a 4-day period, following which the cells appeared to enter a resistant starvation state where cell numbers remained constant over the subsequent 3-4 weeks . This starvation response was induced by carbon or phosphorous limitation, but not by nitrogen limitation in the form of amino acids where cells became non-culturable after 4 days . Amino acid utilization in the absence of a carbon source may be an essential factor for the long-term survival of this bacterium in the stationary phase . Early stationary phase cells showed a greater resistance to oxidative and pH stress compared to 24-h-starved cultures . There was evidence for the formation of a viable but non-culturable state as indicated by a comparison of the numbers of cells with a functional membrane potential (rhodamine 123) against culturable cells on either Todd Hewitt broth agar or sheep blood agar . Long-term survival of S . pyogenes was dependent on both cell wall and protein synthesis, suggesting that starving cultures are a dynamic cell population. Biosci Biotechnol Biochem, 1999 Jun, 63(6), 1107 - 11 Cloning and sequencing of the beta-fructofuranosidase gene from Bacillus sp . V230; Kurimoto M et al.; The beta-fructofuranosidase gene (bff) from Bacillus sp . V230 has been cloned in Escherichia coli and its nucleotide sequence has been analyzed . The product of bff consists of a signal sequence of 32 amino acid (a.a.) residues for secretion and 455 a.a . residues of the extracellular beta-fructofuranosidase . The a.a . sequence of the bff product has similarities with those of the Bacillus subtilis levanscrase (63.7% identity), the Streptococcus mutans fructosyltransferase (33.7%), and the Zymomonas mobilis levanscrase and beta-fructofuranosidase (15%). Biosci Biotechnol Biochem, 1999 Jun, 63(6), 1056 - 62 Cloning, nucleotide sequence, and disruption of Streptococcus mutans glutathione reductase gene (gor); Yamamoto Y et al.; We cloned and sequenced the glutathione reductase gene (gor) of an oxygen-tolerant Streptococcus mutans, and constructed a gor-disruption mutant by homologous recombination . The gor gene consisted of 1,350 bp, coding for a protein of 450 amino acid residues . The deduced amino acid sequence of the S . mutans gor gene product showed extensive similarity with those of glutathione reductases from prokaryotes and eukaryotes . Although the mutant could grow aerobically, it showed no growth in the presence of 2 mM diamide, a thiol-specific oxidant . In contrast, growth of the wild-type strain was not significantly inhibited by 2 mM diamide, and glutathione reductase activity was increased 2.2-fold under these conditions . In addition, the level of glutathione reductase activity in the wild-type strain was increased 3.6-fold upon exposure to air, and the elevated level of the enzyme was retained throughout the aerobic growth . Thus, glutathione reductase may be important in protection of S . mutans against oxidative stress. Nat Med, 1999 Aug, 5(8), 924 - 9 Rapid selection of complement-inhibiting protein variants in group A Streptococcus epidemic waves; Hoe NP et al.; Serotype M1 group A Streptococcus strains cause epidemic waves of human infections long thought to be mono- or pauciclonal . The gene encoding an extracellular group A Streptococcus protein (streptococcal inhibitor of complement) that inhibits human complement was sequenced in 1,132 M1 strains recovered from population-based surveillance of infections in Canada, Finland and the United States . Epidemic waves are composed of strains expressing a remarkably heterogeneous array of variants of streptococcal inhibitor of complement that arise very rapidly by natural selection on mucosal surfaces . Thus, our results enhance the understanding of pathogen population dynamics in epidemic waves and infectious disease reemergence. J Endod, 1999 Apr, 25(4), 239 - 42 Cytotoxic T lymphocytes versus streptococcal colonization in periapical granulomas; Hren NI et al.; Seventeen dental periapical lesions were investigated to study bacterial colonization . Periapical lesions, obtained after apicotomy, were also enzymatically desegregated to quantitatively analyze lymphocyte subpopulations by flow cytometry . Fourteen samples yielded a positive bacterial growth when homogenized and cultured . We isolated enough lymphocytes to make flow cytometric analysis from 12 samples . A significant increase in interleukin-2 receptor and ICAM-1 molecule expression on T cells was found, compared with peripheral blood lymphocytes . Furthermore, a decreased expression of interleukin-2 receptors and HLA DR molecules on CD8+ T cells was found in granulomas predominantly colonized by Streptococcus spp., compared with lesions predominantly colonized by strict anaerobes. Eur J Med Res, 1999 Jul 28, 4(7), 283 - 5 Congenital asplenia detected in a 60 year old patient with septicemia; Germing U et al.; A 60 year-old female who had never been seriously ill, was brought to the emergency ward after she had been found somnolent at home . She developed renal failure, meningitis, respiratory distress and disseminated intravascular coagulation . Overwhelming septicemia was evident, and streptococcus pneumoniae was isolated from blood and cerebrospinal fluid . Surprisingly, peripheral blood smears showed numerous Howell-Jolly-bodies, indicating severe impairment of splenic function . On abdominal ultrasound, CT-scan, and scintigraphy no spleen could be detected . There was no history of abdominal surgery . Apparently, congenital asplenia, which was not noticed until the age of 60, was responsible for the patient's life-threatening septicemia . We suggest that, in cases of severe septicemia, the examination of a blood smear is useful to detect functional (or congenital) asplenia. J Infect, 1999 May, 38(3), 185 - 90 A Streptococcus pyogenes outbreak caused by an unusual serotype of low virulence: the value of typing techniques in outbreak investigations; Davies BI et al.; OBJECTIVES: To investigate and stop the spread of an outbreak of Streptococcus pyogenes infection in a district general hospital, involving 19 patients and two nurses over a 20-day period . METHODS: All suspected persons were investigated using conventional bacteriological techniques, followed by M, T and exotoxin gene-typing of the isolates in a national reference laboratory . RESULTS: 11 patients and both nurses were associated with the acute surgical ward on one floor of the hospital . They were infected with serotype M 9/T B3264, a strain with apparently low virulence, which has not been previously associated with outbreaks . Two other patients on the same floor each had different types and there were two clusters of another S . pyogenes serotype on other floors, each involving two patients . Two (unrelated) patients yielded other types of S . pyogenes . The patients were not seriously ill but had some delay in wound healing . CONCLUSIONS: The value of typing the isolates of S . pyogenes in this outbreak was in defining which patients were involved . No added value could be ascribed to the exotoxin gene-typing results. Chest, 1999 Jul, 116(1), 115 - 9 A randomized, crossover design study of the pharmacology of extended-spectrum fluoroquinolones for pneumococcal infections; Amsden GW et al.; STUDY OBJECTIVES: The objectives of this study were to characterize the single-dose and steady-state plasma pharmacokinetics of IV levofloxacin and IV alatrofloxacin, and to compare the results to pneumococcal isolate sensitivities in order to estimate the clinical efficacy of current community-acquired pneumonia treatment regimens against pneumococcal infections . DESIGN: Two-way, open-label, randomized, crossover study . PARTICIPANTS: Each of 12 healthy volunteer subjects received IV levofloxacin, 500 mg qd for 7 days, and IV alatrofloxacin, 200 mg qd for 7 days . The two regimens were separated by a 2-week washout period . MEASUREMENTS AND RESULTS: Plasma concentration profiles were collected around the first and final doses of both regimens and were assayed for their respective quinolone concentrations . When the peak concentrations for both agents were compared to standard twofold dilution minimum inhibitory concentration (MIC) values for pneumococcal isolates, it was discovered that the breakpoint MIC value at which each compound would no longer achieve a peak plasma concentration/MIC ratio of at least 12:1 was 0.5 mg/L for levofloxacin and 0.25 mg/L for alatrofloxacin . CONCLUSIONS: Based on the MIC that inhibits 90% of isolates of Streptococcus pneumoniae for both of these agents (1.0 to 2.0 mg/L for levofloxacin and 0.125 to 0.25 mg/L for trovafloxacin), our results indicate that although the once-daily regimen of alatrofloxacin appears to be appropriate for this pathogen, a more aggressive regimen may need to be investigated to optimize the clinical and microbiological effects of levofloxacin. Kansenshogaku Zasshi, 1999 Jun, 73(6), 602 - 5 {Three cases of pneumonia due to mixed infection of bacteria and Mycoplasma pneumoniae}; Okimoto N et al.; We reported three cases of pneumonia due to mixed infection of bacteria (2 cases: Streptococcus pneumoniae, 1 case: methicilline-sensitive Stapholococcus aureus) and Mycoplasma pneumoniae . Increased serum antibody titers of M . pneumoniae were noted in all cases . They were a 36-year-old-female with bronchial asthma, a 74-year-old-male with old pulmonary tuberculosis and a 82-year-old-male with chronic bronchitis . All cases had fever, productive cough with purulent sputum and coarse crackle by auscultation . Leukocytosis was noted in 2 cases . Chest X-ray films showed dense consolidation in all cases, 2 cases were cured by administration of cephems and 1 case was cured by administration of carbapenems and minocycline. Kansenshogaku Zasshi, 1999 Jun, 73(6), 584 - 92 {Report of questionnaire survey for methicillin-resistant Staphylococcus aureus and penicillin-resistant Streptococcus pneumoniae in the Kinki District}; Morimoto K et al.; In the Kinki District (Hyogo area, Osaka City area, Osaka Outskirts area, Nara area and Wakayama area), a questionnaire survey of 30 institutions was conducted for methicillin-resistant Staphylococcus aureus (MRSA) and penicillin-resistant Streptococcus pneumoniae (PRSP) . Median number of their bed was 500 ranging 0 to 1076, 3239 (56%) of the 5815 strains of S . aureus were methicillin-resistant . Although no different prevalence was found among the areas, they were predominantly isolated from sputum of inpatients more than from outpatients, 336 (44%) of the 763 strains of S . pneumoniae were penicillin-resistant . The prevalence rate was the highest in the outpatients in Osaka outskirts . Almost all strains of MRSA were sensitive to vancomycin (VCM) and sulfamethoxazole trimethoprim (ST) . Resistant strains were observed in 2% against arbekacin, 6% against amikacin, 44% against minocycline (MINO), and in 54% against gentamicin (GM) . Almost all strains of PRSP were sensitive to VCM and ST . Resistant strains were observed in 75% against erythromycin, 50% against MINO, and 75% against GM . This survey identified the trend of hospital infection for MRSA and community infection for PRSP, and sensitive drugs for the treatment. J Biochem (Tokyo), 1999 Aug, 126(2), 287 - 95 Structure and enzymatic properties of genetically truncated forms of the water-insoluble glucan-synthesizing glucosyltransferase from Streptococcus sobrinus; Konishi N et al.; Glucosyltransferase-I (GTF-I: 175 kDa) of a cariogenic bacterium, Streptococcus sobrinus 6715, mediates the conversion of water-soluble dextran (alpha-1,6-glucan) into a water-insoluble form by making numerous alpha-1,3-glucan branches along the dextran chains with sucrose as the glucosyl donor . The structures and catalytic properties were compared for two GTF-I fragments, GTF-I' (138 kDa) and GS (110 kDa) . Both lack the N-terminal 84 residues of GTF-I . While GTF-I' still contains four of the six C-terminal repeats characteristic of streptococcal glucosyltransferases, GS lacks all of them . Electron microscopy of negatively stained samples indicated a double-domain structure for GTF-I', consisting of a spherical head with a smaller spherical tail, which was occasionally seen as a long extension . GS was seen just as the head portion of GTF-I' . In the absence of dextran, both fragments simply hydrolyzed sucrose with similar K(m) and k(cat) values at low concentrations (<5 mM) . At higher sucrose concentrations (>10 mM), however, GTF-I' exhibited glucosyl transfer activity to form insoluble alpha-1, 3-glucans . So did GS, but less efficiently . Dextran increased the rate and efficiency of the glucosyl transfer by GTF-I' . On removal of the C-terminal repeats of GTF-I' by mild trypsin treatment, this dextran-stimulated transfer was completely lost and the dextran-independent transfer became less efficient . These results indicate that the N-terminal two-thirds of the GTF-I sequence are organized as a structurally and functionally independent domain to catalyze not only sucrose hydrolysis but also glucosyl transfer to form alpha-1,3-glucan chains, although not efficiently; the C-terminal repeat increases the efficiency of the intrinsic glucosyl transfer by the N-terminal domain as well as rendering the whole molecule primer-dependent for far more efficient insoluble glucan synthesis. Int J Antimicrob Agents, 1999 Jul, 12(2), 107 - 14 Comparative efficacies of levofloxacin and ciprofloxacin against Streptococcus pneumoniae in a mouse model of experimental septicaemia; Onyeji CO et al.; The in vivo efficacies of levofloxacin and ciprofloxacin were compared against three clinical isolates of Streptococcus pneumoniae, using a mouse protection model . Two strains (SP 22 and SP 28) were penicillin-sensitive while one strain (SP 46) was penicillin-resistant . Each strain had identical susceptibility to both drugs . Using mice with renal impairment induced by uranyl nitrate injection, the elimination half-life of each antibiotic was prolonged to approximate human pharmacokinetic profiles of the drugs . The dosing regimen of each drug that yielded serum levels in mice which mimic human therapeutic concentrations of the drugs, were designed . One hour after intraperitoneal inoculation with minimum lethal dose of each strain, either levofloxacin at a dosing regimen of 10.6 mg/kg every 8 h or ciprofloxacin at 9.5 mg/kg every 8 h was subcutaneously administered for a total of six or 15 doses . In treatment, monitored daily for 5-8 days, levofloxacin resulted in higher survival compared with ciprofloxacin for the three strains . For example, percent survival following levofloxacin treatment recorded at day 4 postinfection with SP 22, SP 28 and SP 46 were 41, 90 and 30%, respectively, while the corresponding values after ciprofloxacin treatment were 27, 75 and 16%, respectively . However, statistical analysis did not reveal a significant difference (p > 0.05) . The lack of significant difference observed in the efficacies of both drugs reflected the comparability of their 24-h AUC/MIC ratios . It is suggested that, with some strains of S . pneumoniae, the efficacy of levofloxacin may be equivalent to that of ciprofloxacin in the treatment of systemic pneumococcal infections caused by susceptible strains of the organism. J Acquir Immune Defic Syndr, 1999 Jul 1, 21(3), 194 - 202 Bacterial vaginosis-associated microflora isolated from the female genital tract activates HIV-1 expression; Al-Harthi L et al.; Alteration of cervicovaginal microbial flora can lead to vaginosis, which is associated with an increased risk of HIV-1 transmission . We recently characterized a soluble HIV-inducing factor (HIF) from the cervicovaginal lavage (CVL) samples of women . The goals of this study were to determine the effect of cervicovaginal microflora on HIV-1 expression and to elucidate the relationship between HIF activity and microflora . Physiologically relevant microorganisms, Mycoplasma, diphtheroid-like bacteria, Gardnerella vaginalis, Streptococcus agalactiae, and Streptococcus constellatus, cultured from the CVL of a representative woman with a clinical condition of bacterial vaginosis and possessing HIF activity, induced HIV-1 expression . The magnitude of virus induction varied widely with the greatest stimulation induced by diphtheroid-like bacteria and Mycoplasma . The transcriptional induction by Mycoplasma was mediated by activation of the KB enhancer, an activation mechanism shared with HIF . Also as with HIF, Mycoplasma induced AP-1 dependent transcription . Polymerase chain reaction (PCR)-based speciation showed that the isolate was M . hominis . Our data indicate that bacterial vaginosis-associated microflora can enhance HIV-1 transcription and replication and identify M . hominis as a potential source for HIF activity . The virus-enhancing activities associated with the microflora and HIF may increase genital tract viral load, potentially contributing to HIV transmission. Eur J Clin Microbiol Infect Dis, 1999 May, 18(5), 376 - 9 Resistance patterns of Streptococcus pneumoniae from children in central Italy; Ronchetti MP et al.; Nasopharyngeal swabs were collected from children aged 3-5 years in central Italy who were attending day-care centres or hospital outpatient clinics . One hundred and twenty-one strains of Streptococcus pneumoniae isolated were tested for susceptibility to penicillin, cefotaxime, erythromycin, clindamycin, tetracycline, chloramphenicol and cotrimoxazole . A high prevalence of penicillin-resistant (14%), erythromycin-resistant (60%) and multiply resistant strains (53%) were found . An unusual finding was that 49 of the 64 (76.6%) multiply resistant strains were penicillin-susceptible, 28 serogroup 6 strains also being resistant to the other antibiotics tested . Such strains have not previously been reported from Italy but have the same features as strains recently found in child carriers in the eastern Mediterranean area. Eur J Clin Microbiol Infect Dis, 1999 May, 18(5), 365 - 7 Bacteremia complicated by vertebral osteomyelitis due to Streptococcus bovis; Hechmann Wittrup I et al.; The diagnosis of vertebral osteomyelitis is easily missed, especially in the elderly in whom clinical signs of bacteremia might not be manifest . Spontaneously occurring disc-space infection in adults often has an insidious presentation . The infecting microorganism can be difficult to identify . Although discitis due to Streptococcus bovis is occasionally found, it is often difficult to fully confirm the diagnosis . Here, a case of vertebral osteomyelitis due to this microorganism is reported. Rev Med Univ Navarra, 1998 Jan-Mar, 42(1), 14 - 7 {Chlamydia pneumoniae pneumonia}; Latorre G et al.; Chlamydia pneumoniae causes respiratory tract infections, and it is transmitted by air and fomites . It is probably more frequent than it is described, due to asymptomatic or mild symptomatic patients . They respond to macrolides, tetracyclines and quinolones, though patients may recover slowly . An increase of the incidence of pneumonia, caused by Chlamydia pneumoniae, is shown in recent multicenter surveys, being even more frequent than Streptococcus pneumoniae and Mycoplasma pneumoniae . Recently it has been demonstrated an association between coronary artery disease and atherosclerosis with Chlamydia pneumoniae infection . Special attention must be paid to the cardiovascular complications of Chlamydia pneumoniae . We describe six clinical cases of Chlamydia pneumoniae pneumonia in which two of them suffered from ischemic artery disease as a complication of the infection. Ocul Immunol Inflamm, 1999 Jun, 7(2), 69 - 74 Behçet's disease and antibody titers to various heat-shock protein 60s; Tanaka T et al.; It has been suggested that the 65 kDa heat-shock protein (HSP) of Streptococcus in recurrent aphthae within the oral cavity may be involved in the uveoretinitis of Behcet's disease, possibly through sensitization of the immune system . To investigate this possibility, we examined serum antibody titers for various members of the 60 kDa family of HSPs and their implications with regard to a role for HSP60s in Behcet's disease . We isolated HSP60 of Streptococcus pyogenes from the margin of oral aphthae in one Behcet's disease patient with severe uveoretinitis and the HSP60s of Yersinia enterocolitica, retinoblastoma cell line clone Y79, and bovine retinal extract and investigated the reaction of each of these HSP60s with 100-fold diluted serum samples from 20 Behcet's disease patients using anti-HSP60 antibody titers determined by ELISA . The anti- Streptococcus HSP60 antibody and anti-retinal HSP60 antibody titers of the 100-fold diluted serum samples from the Behcet's disease patients were both significantly higher than those of similarly diluted serum samples from healthy donors . The results of the ELISA antibody titer assay showed that, although the various HSP60s share a common basic antigenicity, they differed in reactivity to the anti-HSP60 antibodies in the sera of the Behcet's disease patients . The results indicate that subtle but significant differences exist in the antigenicity of the various HSP60s tested, all of which share a common basic antigenicity and are of approximately the same molecular weight, and suggest that an immuno-cross-reaction between retinal and streptococcal HSPs and a related autoimmune response may be involved in the development of Behcet's disease. Protein Expr Purif, 1999 Jul, 16(2), 331 - 9 Penicillin-binding protein 2a of Streptococcus pneumoniae: expression in Escherichia coli and purification and refolding of inclusion bodies into a soluble and enzymatically active enzyme; Zhao G et al.; Penicillin-binding proteins (PBPs), targets of beta-lactam antibiotics, are membrane-bound enzymes essential for the biosynthesis of the bacterial cell wall . PBPs possess transpeptidase and transglycosylase activities responsible for the final steps of the bacterial cell wall cross-linking and polymerization, respectively . To facilitate our structural studies of PBPs, we constructed a 5'-truncated version (lacking bp from 1 to 231 encoding the N-terminal part of the protein including the transmembrane domain) of the pbp2a gene of Streptococcus pneumoniae and expressed the truncated gene product as a GST fusion protein in Escherichia coli . This GST fusion form of PBP2a, designated GST-PBP2a*, was expressed almost exclusively as inclusion bodies . Using a combination of high- and low-speed centrifugation, large amounts of purified inclusion bodies were obtained . These purified inclusion bodies were refolded into a soluble and enzymatically active enzyme using a single-step refolding method consisting of solubilization of the inclusion bodies with urea and direct dialysis of the solubilized preparations . Using these purification and refolding methods, approximately 37 mg of soluble GST-PBP2a* protein was obtained from 1 liter of culture . The identity of this refolded PBP2a* protein was confirmed by N-terminal sequencing . The refolded PBP2a*, with or without the GST-tag, was found to bind to BOCILLIN FL, a beta-lactam, and to hydrolyze S2d, an analog of the bacterial cell wall stem peptides . The S2d hydrolysis activity of PBP2a* was inhibited by penicillin G . In conclusion, using this expression system, and the purification and refolding methods, large amounts of the soluble GST-PBP2a* protein were obtained and shown to be enzymatically active . Int J Antimicrob Agents, 1999 Jul, 12(2), 141 - 4 In vitro activities of the oxazolidinone compounds linezolid (PNU-100766) and eperzolid (PNU-100592) against middle ear isolates of Streptococcus pneumoniae; Kearney JA et al.; Two hundred and sixteen isolates of Streptococcus pneumoniae recovered between 1994 and 1996 from the middle ears of children with acute otitis media were tested for their susceptibility to penicillin, erythromycin, clindamycin and the oxazolidinones, linezolid (PNU-100766) and eperezolid (PNU-100592) . There were 116 isolates from the Children's Hospital of Pittsburgh and 100 isolates from a national collection . Eighty percent of the local strains were susceptible to penicillin (MIC < 0.1 mg/l); 20% of the local strains and all of the national strains had reduced susceptibility to penicillin . All strains of S . pneumoniae tested had an MIC < 2.0 mg/l for both oxazolidinones . A regional difference was noted in the frequency of resistance to erythromycin with local isolates being more susceptible than isolates from the national collection . This difference was most pronounced among the high-level penicillin-resistant strains of S . pneumoniae. FEMS Microbiol Lett, 1999 Jul 1, 176(1), 91 - 6 Identification of lactoferrin-binding proteins in bovine mastitis-causing Streptococcus uberis; Fang W et al.; All strains of Streptococcus uberis evaluated bound to lactoferrin (Lf) in milk as detected by polyacrylamide gel electrophoresis and Western blotting . A biotin-avidin-based microplate binding assay and ELISA also revealed that these bacterial strains bound to purified Lf . Binding of bacteria of Lf was not inhibited by mannose and galactose, indicating that glycosidic domains of the Lf molecule were not involved in binding . Lf binding was also unaffected by bovine transferrin . Western blot analysis demonstrated that there were at least two bacterial proteins involved in Lf-binding . Lf binding by S . uberis could enable this bacterium to acquire iron necessary for its growth. Rev Rhum Engl Ed, 1999 Jun, 66(6), 344 - 6 Pneumococcal polyarticular septic arthritis in a patient with rheumatoid arthritis; Lohse A et al.; Rheumatoid arthritis is the most commonly reported host-related risk factor for septic arthritis . This risk is highest in severe, seropositive, long-standing (mean, 10 years) rheumatoid arthritis responsible for extraarticular symptoms and treated with systemic glucocorticoids . The clinical presentation of the joint infection is often atypical, leading to diagnostic wanderings . In 25% of cases, the infection is polyarticular, with 3.5 involved joints on average . Staphylococcus aureus is the most common causative organism . Streptococcus pneumoniae causes 5% of all cases of septic arthritis and is more often responsible for polyarticular infections than other organisms . Polyarticular septic arthritis carries a poor prognosis, with a mortality rate of 50% in rheumatoid arthritis patients . Despite its low incidence, polyarticular septic arthritis should be routinely considered in the differential diagnosis of rheumatoid flares . We report a case of pneumococcal septic arthritis involving five joints in a patient with known rheumatoid arthritis . Three other cases with involvement of more than four joints have been published. Pathol Biol (Paris), 1999 May, 47(5), 519 - 25 {Selection of virulent mutants of Streptococcus pneumoniae . Utilization of a murine model of septicemia}; Amory-Rivier C et al.; Genetic construction of virulence deficient mutant is a strategy to analyse virulence genes of Streptococcus pneumoniae and was used to virulence factors as capsule, pneumolysin, autolysin and PspA . We perform a model allowing the in vivo positive selection of virulent S . pneumoniae mutants . Mice which are the most susceptible animals to pneumococcal infection, offer the best model for screening virulent S . pneumoniae . Indeed, after intraperitoneal injection of bacterial mix which was composed to a lot of avirulent bacteria (6 log10 CFU per mouse) (V1015 strain, DL50 = 7.05) and few virulent pneumococci (1 to 2 log10 CFU per mouse) (P4241 strain, DL50 < 1), mice cleared all avirulent bacteria but not virulent pneumococci . Thus, mice dead in 3 to 4 days with septicaemia and positive hemoculture contained only virulent strain . This model was validated by in vivo selection of a virulent mutant (V1042, DL50 = 4.1) which was obtained after transformation of avirulent strain V1015 with the genomic fragment of virulent strain P4241 . Our model of screening was the only one allowing detection of virulent S . pneumoniae mutants . This new genetic strategy which consisted in gene addition and used mouse as selection agent, could be used to discover new virulence genes required to in vivo bacterial development. Pathol Biol (Paris), 1999 May, 47(5), 478 - 82 {Nasopharyngeal carriage of resistance pneumococci in day-care centers in the Alps-Maritimes region}; Pradier C et al.; To provide ongoing information on regional trends of antibiotic resistance prevalence to pneumococci, a cross selectional survey was conducted on a large representative sample of children attending day-care centers . Children were analyzed in spring (n = 378) and autumn (n = 379) for nasopharyngeal carriage . Streptococcus pneumoniae was detected in 149 children (39.4%) in Spring and 204 (59.8%) in Autumn . Half of these isolated strains showed penicillin insensitivity or resistance . A high proportion of children (43.6% in spring and 47.5% in autumn) had been treated with antibiotics during the 3 months prior to sample collection; 21.6% of isolated strains were serotype 6B, 20.1% type 23F, 18.9% type 19A and 19F, 11.5% type 14 . Reduced susceptibility was frequently noted in serotype 23F, 14 and 19F, representing 93%, 94% and 46% of identified serotypes, respectively . Acquisition of a strain of PRP was correlated with prescription of antibiotics during the previous three months (p < 0.05) . This type of survey on children in day-care centers can contribute to the understanding of regional variations in antibiotic resistance and provide information for epidemiological surveillance. Pathol Biol (Paris), 1999 May, 47(5), 422 - 9 {Severe community-acquired bacterial pneumonia from Streptococcus pneumonia in HIV-infected patients: epidemiology and prognostic features of mortality}; Megarbane B et al.; In HIV-infected patients, community-acquired pneumonia due to Streptococcus pneumoniae is more common, more often associated with bacteremia, and more likely to recur . The epidemiology of this condition is discussed based on a literature review . Factors predictive of mortality in severe cases managed in intensive care units are analyzed. Infect Immun, 1999 Aug, 67(8), 4128 - 33 Intranasal immunization with pneumococcal polysaccharide conjugate vaccines protects mice against invasive pneumococcal infections; Jakobsen H et al.; Host defenses against Streptococcus pneumoniae depend largely on opsonophagocytosis mediated by antibodies and complement . Since pneumococcus is a respiratory pathogen, mucosal immune responses may play a significant role in the defense against pneumococcal infections . Thus, mucosal vaccination may be an alternative approach to current immunization strategies, but effective adjuvants are required . Protein antigens induce significant mucosal immunoglobulin A (IgA) and systemic IgG responses when administered intranasally (i . n.) with the glyceride-polysorbate based adjuvant RhinoVax (RV) both in experimental animals and humans . The immunogenicity and efficacy of pneumococcal polysaccharide conjugate vaccines (PNC) of serotypes 1 and 3 was studied in mice after i.n . immunization with RV . Antibodies were measured in serum (IgM, IgG, and IgA) and saliva (IgA) and compared to antibody titers induced by parenteral immunization . The PNCs induced significant systemic IgG and IgA antibodies after i.n . immunization only when given with RV and, for serotype 1, serum IgG titers were comparable to titers induced by subcutaneous immunization . In addition, i.n . immunization with PNC-1 in RV elicited detectable mucosal IgA . These results demonstrate that RV is a potent mucosal adjuvant for polysaccharides conjugated to proteins . A majority of the PNC-1-immunized mice were protected against both bacteremia and pneumonia after i.n . challenge with a lethal dose of serotype 1 pneumococci, and protection correlated significantly with the serum IgG titers . Similarly, the survival of mice immunized i.n . with PNC-3 in RV was significantly prolonged . These results indicate that mucosal vaccination with PNC and adjuvants may be an alternative strategy for prevention against pneumococcal infections. Infect Immun, 1999 Aug, 67(8), 4119 - 27 Molecular and functional characteristics of a protective human monoclonal antibody to serotype 8 Streptococcus pneumoniae capsular polysaccharide; Zhong Z et al.; The structural characteristics and biological activity of human antibodies that are reactive with the capsular polysaccharides of most serotypes of Streptococcus pneumoniae, including serotype 8, are unknown . This paper describes the generation, molecular structure, and protective efficacy of a human monoclonal antibody (MAb) reactive with the capsular polysaccharide of serotype 8 Streptococcus pneumoniae . We generated the immunoglobulin M(kappa) {IgM(kappa)} MAb D11 by Epstein-Barr virus transformation of peripheral lymphocytes from a Pneumovax recipient . Nucleic acid sequence analysis revealed that MAb D11 uses V3-15/V(H)3 and A20/V(kappa) gene segments with evidence of somatic mutation . In vitro studies revealed MAb D11-dependent complement deposition on the capsule of serotype 8 organisms via either the classical or the alternative complement pathway . In vivo, MAb D11 prolonged the survival of both normal and C4-deficient mice with lethal serotype 8 S . pneumoniae infection . Our findings demonstrate that a serotype-specific human IgM with certain structural and functional characteristics was protective in mice lacking a functional classical complement pathway and show that alternative complement pathway activation is an important determinant of pneumococcal protection. Infect Immun, 1999 Aug, 67(8), 3780 - 5 Secretion of functional salivary peptide by Streptococcus gordonii which inhibits fimbria-mediated adhesion of Porphyromonas gingivalis; Kataoka K et al.; Porphyromonas gingivalis, a putative periodontopathogen, can bind to human salivary components with its fimbriae . We have previously shown that fimbriae specifically bind to a peptide domain shared by a major salivary component, i.e., proline-rich (glyco)proteins (PRPs) . The synthetic domain peptide PRP-C (pPRP-C) significantly inhibits the fimbrial binding to PRPs . In this study, a recombinant strain of Streptococcus gordonii secreting pPRP-C was generated as a model of a possible approach to prevent the oral colonization by the pathogen . A duplicate DNA fragment (prpC) encoding pPRP-C was obtained by self-complementary annealing of synthetic oligonucleotides . prpC was connected downstream to a promoter and a gene encoding a signal peptide of Streptococcus downei glucosyltransferase I in frame . The linked fragments were inserted into the plasmid pMNK-4 derived from pVA838 . The constructed plasmid was inserted to produce the transformant S . gordonii G9B, which then successfully secreted recombinant pPRP-C (r-pPRP-C) of the expected size . The concentrated bacterial culture supernatant containing r-pPRP-C inhibited the binding of P . gingivalis cells and fimbriae to PRP1 in a dose-dependent manner up to 72 and 77%, respectively . The r-pPRP-C concentrate also inhibited the coaggregation of P . gingivalis with various streptococcal strains as effectively as synthetic pPRP-C in a dose-dependent manner . Collectively, pPRP-C was found to be able to prevent P . gingivalis adherence to salivary receptor protein and plaque-forming bacteria . These results suggest that this recombination approach with a nonperiodontopathic bacterium may be suitable for the therapeutic prevention of P . gingivalis adherence to the oral cavity. Infect Immun, 1999 Aug, 67(8), 3750 - 6 Pneumolysin, a protein toxin of Streptococcus pneumoniae, induces nitric oxide production from macrophages; Braun JS et al.; Nitric oxide (NO) production by inducible NO synthase (iNOS) during inflammation is an essential element of antimicrobial immunity but can also contribute to host-induced tissue damage . Under conditions of bacterial sepsis, large amounts of NO are produced, causing hypotension, a critical pathological feature of septic shock . In sepsis caused by gram-positive organisms, the bacterial factors contributing to host NO production are poorly characterized . We show that a soluble toxin of Streptococcus pneumoniae, pneumolysin (Pln), is a key component initiating NO production from macrophages . In contrast to wild-type bacteria, a mutant of S . pneumoniae lacking Pln failed to elicit NO production from murine macrophages . Purified recombinant Pln induced NO production at low concentrations and independently of exogenous gamma interferon (IFN-gamma) priming of RAW 264.7 macrophages . However, IFN-gamma was essential for Pln-induced NO production, since primary macrophages from mice lacking the IFN-gamma receptor or interferon regulatory factor 1, a transcription factor essential for iNOS expression, failed to produce NO when stimulated with Pln . In addition, Pln acts as an agonist of tumor necrosis factor alpha and interleukin 6 production in macrophages . The properties of Pln, previously identified as a pore-forming hemolysin, also include a role as a general inflammatory agonist. J Dairy Sci, 1999 Jul, 82(7), 1465 - 81 Effect of bovine somatotropin on neutrophil functions and clinical symptoms during Streptococcus uberis mastitis; Hoeben D et al.; The effect of recombinant bovine somatotropin (bST) on the chemiluminescence, diapedesis, and expression of adhesion receptors (CD11a, CD11b, CD18) of isolated polymorphonuclear leukocytes was studied . The plasma concentrations of insulin-like growth factor-I (IGF-I), bST, cortisol, and alpha-lactalbumin were also monitored . In addition, general and local clinical symptoms and the differentiation of circulating leukocytes were also studied during experimentally induced Streptococcus uberis mastitis in cows . Ten cows were infected with 500 cfu of S . uberis O140J in both left quarters . Five cows were subcutaneously treated with 500 mg of recombinant bST 7 d before and after infection, and 5 control cows received the excipient . General (fever, tachycardia, inappetance, and depression) and local symptoms (swelling, pain, firmness, and flecks in milk) were more acute, severe, and longer-lasting in control cows . Treatment with bST had no effect on chemiluminescence and diapedesis of circulating polymorphonuclear leukocytes and no effect on the expression of adhesion receptors . Recombinant bST induced significantly higher IGF-I and bST concentrations in plasma . The leukopenia observed after infection was less pronounced in the bST-treated cows, and the number of circulating band neutrophils and metamyelocytes was significantly lower in the treated group . The concentration of cortisol did not differ between both groups, but the blood concentration of alpha-lactalbumin significantly increased in both groups from 6 d after infection . These results showed that treatment with recombinant bST improves animal welfare by protecting the cows from severe local and general clinical symptoms during subsequent S . uberis mastitis, but that it has no effect on chemiluminescence, diapedesis, and the expression of adhesion receptors of circulating polymorphonuclear leukocytes. N Engl J Med, 1999 Jul 22, 341(4), 233 - 9 Decreased susceptibility of Streptococcus pneumoniae to fluoroquinolones in Canada . Canadian Bacterial Surveillance Network; Chen DK et al.; BACKGROUND: Fluoroquinolones are now recommended for the treatment of respiratory tract infections due to Streptococcus pneumoniae, particularly when the isolates are resistant to beta-lactam antibiotics . Although pneumococci with reduced susceptibility to fluoroquinolones have been identified, their prevalence has not been determined in a defined population . METHODS: We performed susceptibility testing on 7551 isolates of S . pneumoniae obtained from surveillance in Canada in 1988 and from 1993 to 1998 . Pneumococci with reduced susceptibility to fluoroquinolones (defined as a minimal inhibitory concentration of ciprofloxacin of at least 4 microg per milliliter) were further characterized . We also examined antibiotic prescriptions dispensed in Canadian retail pharmacies . RESULTS: Between 1988 and 1997, fluoroquinolone prescriptions increased from 0.8 to 5.5 per 100 persons per year . The prevalence of pneumococci with reduced susceptibility to fluoroquinolones increased from 0 percent in 1993 to 1.7 percent in 1997 and 1998 (P=0.01) . Among adults, the prevalence increased from 1.5 percent in 1993 and 1994 combined to 2.9 percent in 1997 and 1998 combined . The prevalence was higher in isolates from older patients (2.6 percent among those 65 years of age or older vs . 1.0 percent among those 15 to 64 years of age, P<0.001) and among those from Ontario (1.5 percent, vs . 0.4 percent among those from the rest of Canada; P< 0.001) . Fluoroquinolone use was greatest among the elderly and in Ontario . The 75 isolates (17 serotypes) of pneumococci with reduced susceptibility to fluoroquinolones were submitted by 40 laboratories in eight provinces . Reduced susceptibility to fluoroquinolones was associated with resistance to penicillin . CONCLUSIONS: The prevalence of pneumococci with reduced susceptibility to fluoroquinolones is increasing in Canada, probably as a result of selective pressure from the increased use of fluoroquinolones. South Med J, 1999 Jul, 92(7), 728 - 31 Bacterial complications of strongyloidiasis: Streptococcus bovis meningitis; Link K et al.; We report the case of a 64-year-old veteran who had Streptococcus bovis meningitis as a result of a long latent Strongyloides infection that became acute when he was treated with prednisone . We reviewed 38 reported cases of serious bacterial infections associated with strongyloidiasis . Patients most frequently had nonspecific gastrointestinal symptoms . Of these 38 patients, 21 (55%) had meningitis, and 28 (73%) had bacteremia that was polymicrobial in 3 cases (8%) . Other sites of infection included lung, bone marrow, ascites, mitral valve, and lymph node . Most infections were due to enteric gram-negative bacteria . There is one previously reported case of S bovis meningitis . Thirty-four of the patients (89%) were immunosuppressed; 21 of these (55%) were taking pharmacologic doses of adrenal corticosteroids . Thirty-three of the 38 (87%) patients died . Patients with enteric bacterial infection without an obvious cause should be tested for the presence of strongyloidiasis. Vet Microbiol, 1999 Jun 15, 67(2), 143 - 57 Detection of virulent strains of Streptococcus suis type 2 and highly virulent strains of Streptococcus suis type 1 in tonsillar specimens of pigs by PCR; Wisselink HJ et al.; We developed a PCR assay for the rapid and sensitive detection of virulent Streptococcus suis type 2 and highly virulent S . suis type 1 in tonsillar specimens from pigs . The PCR primers were based on the sequence of the gene encoding the EF-protein of virulent S . suis type 2 strains (MRP+EF+) and highly virulent S . suis type 1 strains (MRP(s)EF+) and of the EF protein of weakly virulent S . suis type 2 strains (MRP+EF) . The latter strains give rise to larger PCR products than the virulent strains of S . suis type 1 and 2 . A positive control template was included in the assay to identify false negative results . The PCR was evaluated using tonsillar specimens from herds known (or suspected) to be infected and herds without an S . suis history . The results obtained with the PCR assay were compared with the results obtained with a newly developed bacteriological examination . In this bacteriological examination we were able to identify the EF-positive strains directly in the tonsillar specimens . From the 99 tonsils examined, 48 were positive in the PCR and 51 negative . All specimens from which EF-positive S . suis strains were isolated were also positive in the PCR assay . Three samples were positive in the PCR, but negative by bacteriological examination . The results demonstrated that the PCR is a highly specific and sensitive diagnostic tool for the detection of pigs carrying virulent strains of S . suis type 2 and highly virulent strains of type 1 . Application of the assay may contribute to the control of S . suis infections. WMJ, 1999 May-Jun, 98(3), 42 - 5 Prevalence of penicillin resistant and multi-drug resistant Streptococcus pneumoniae at a children's hospital; Kehl SC et al.; Streptococcus pneumoniae isolated at Children's Hospital of Wisconsin during the winter of 1994 to 1995 and the winter of 1996 to 1997 were tested for susceptibility to penicillin, cephalosporins and other potentially therapeutically useful antimicrobial agents to determine the prevalence of penicillin and multi-drug resistant isolates . During those years, the prevalence of S . pneumomiae not susceptible to penicillin was 27% and 28%, respectively, with 14% and 18%, respectively, of the respiratory isolates being high-level penicillin resistant . Despite the stable numbers of penicillin resistant isolates, there was evidence of significant increase in the resistance of these isolates to other antimicrobial agents . Respiratory isolates not susceptible to cefotaxime increased (p = .01; Fisher exact test) from 3% in 1995 to 20% in 1997 . There was also a significant increase in the isolates not susceptible to erythromycin (p = .09; Fisher exact test) and trimethoprim/sulfamethoxazole (p < .01; Fisher exact test) . This increase in resistance to multiple antimicrobial agents has significant implications for antibiotic therapy of children with infections likely to be due to Streptococcus pneumoniae. Mil Med, 1999 Jul, 164(7), 471 - 4 Self-collection of group B Streptococcus cultures in pregnant women; Spieker MR et al.; OBJECTIVE: This study assesses the sensitivity of self-collected rectovaginal culture specimens for group B Streptococcus by pregnant patients . METHODS: A volunteer sample of 240 pregnant women at 28 weeks gestation self-collected rectovaginal culture swabs to screen for the presence of group B Streptococcus . The patients' physicians collected second specimens for comparison . RESULTS: Twenty-four of 240 women grew group B Streptococcus on at least one culture (incidence, 10%) . Twenty physician-collected specimens and 19 patient-collected specimens were positive (83 and 79% sensitivity, respectively) . Fifteen patients (62.5%) had both physician-collected and patient-collected cultures grow group B Streptococcus . Cohen's kappa (kappa = 0.75) indicates a high degree of agreement between patient-collected and physician-collected cultures . CONCLUSIONS: Pregnant women are as likely as their attending physicians to obtain positive cultures for group B Streptococcus by self-collection of rectovaginal swabs. Plasmid, 1999 Jul, 42(1), 67 - 72 Streptococcal reporter gene-fusion vector for identification of in vivo expressed genes; Kili AO et al.; To study streptococcal genes that are specifically induced in the host during endocarditis, we have developed a novel plasmid for use in in vivo expression technology (IVET) . This IVET uses an integration plasmid, pAK36, that carries dual (amy-cat) reporter genes . A gene-fusion strain library was constructed with the plasmid randomly inserted into the chromosome of Streptococcus gordonii V288 by insertion-duplication . The library was inoculated intravenously into a rabbit that had been prepared for experimental endocarditis . Beginning 6 h after the inoculation, the rabbit was given chloramphenicol (Cm) intravenously twice a day to a final serum level of 5 microg/ml and was euthanized 3 days later . The aortic valve vegetations containing Cm(R) S . gordonii clones were cultured . Colonies were screened in vitro for negative amylase activity and sensitivity to Cm . Forty-eight such colonies showed 13 different insertion patterns when Southern hybridization blots were probed with labeled pAK36 . For each of the 13 isolates, the gene fragment proximal to the insertion of the reporter amy-cat was cloned, and its nucleotide sequence was determined . Functions of these genes were inferred by their homology to known genes . Therefore, this novel IVET vector can be useful for identification of in vivo induced genes in S . gordonii and other streptococcal species . J Clin Periodontol, 1999 Jul, 26(7), 469 - 73 Bacteremia after dental treatment in mentally handicapped people; Messini M et al.; Bacteremia may occur after disruption of the oral mucous membrane, particularly after dental treatment . 18 mentally handicapped patients who underwent dental treatment with general anesthesia were included in our study . None of the patients had general illnesses or received antibiotic protection . From each patient several blood samples were drawn aseptically during dental treatment and cultured . The majority of aerobic bacteria recovered belonged to Streptococcus sp and Gemella sp., anaerobic bacteria mainly belonged to Porphyromonas gingivalis and Peptostreptococcus sp . Resistance of the isolated bacteria to penicillin as well as to oxacillin, erythromycin and Co-trimoxazole was substantial . The highest resistance rate could be shown against fucidic acid. Mol Microbiol, 1999 Jul, 33(1), 208 - 19 The R28 protein of Streptococcus pyogenes is related to several group B streptococcal surface proteins, confers protective immunity and promotes binding to human epithelial cells; Stalhammar-Carlemalm M et al.; The R28 protein is a surface molecule expressed by some strains of Streptococcus pyogenes (group A streptococcus) . Here, we present evidence that R28 may play an important role in virulence . Sequence analysis demonstrated that R28 has an extremely repetitive sequence and can be viewed as a chimera derived from the three surface proteins Rib, alpha and beta of the group B streptococcus (GBS) . Thus, the gene encoding R28 may have originated in GBS . The R28 protein promotes adhesion to human epithelial cells, as shown by experiments with an R28-negative mutant and by the demonstration that antibodies to highly purified R28 inhibited adhesion . In a mouse model of lethal intraperitoneal S . pyogenes infection, antibodies to R28 conferred protective immunity . However, the virulence of an R28-negative mutant was similar to that of the parental strain in the intraperitoneal infection model . Together, these data indicate that R28 represents a novel type of adhesin expressed by S . pyogenes and that R28 may also act as a target for protective antibodies at later stages of an infection . We consider the hypothesis that R28 played a pathogenetic role in the well-known epidemics of childbed fever (puerperal fever), which were caused by S . pyogenes . A role for R28 in these epidemics is suggested by epidemiological data. Mol Microbiol, 1999 Jul, 33(1), 128 - 38 The molecular characterization of the first autolytic lysozyme of Streptococcus pneumoniae reveals evolutionary mobile domains; Garcia P et al.; A biochemical approach to identify proteins with high affinity for choline-containing pneumococcal cell walls has allowed the localization, cloning and sequencing of a gene (lytC ) coding for a protein that degrades the cell walls of Streptococcus pneumoniae . The lytC gene is 1506 bp long and encodes a protein (LytC) of 501 amino acid residues with a predicted M r of 58 682 . LytC has a cleavable signal peptide, as demonstrated when the mature protein (about 55 kDa) was purified from S . pneumoniae . Biochemical analyses of the pure, mature protein proved that LytC is a lysozyme . Combined cell fractionation and Western blot analysis showed that the unprocessed, primary product of the lytC gene is located in the pneumococcal cytoplasm whereas the processed, active form of LytC is tightly bound to the cell envelope . In vivo experiments demonstrated that this lysozyme behaves as a pneumococcal autolytic enzyme at 30 degrees C . The DNA region encoding the 253 C-terminal amino acid residues of LytC has been cloned and expressed in Escherichia coli . The truncated protein exhibits a low, but significant, choline-independent lysozyme activity, which suggests that this polypeptide adopts an active conformation . Self-alignment of the N-terminal part of the deduced amino acid sequence of LytC revealed the presence of 11 repeated motifs . These results strongly suggest that the lysozyme reported here has changed the general building plan characteristic of the choline-binding proteins of S . pneumoniae and its bacteriophages, i.e . the choline-binding domain and the catalytic domain are located, respectively, at the N-terminal and the C-terminal moieties of LytC . This work illustrates the natural versatility exhibited by the pneumococcal genes coding for choline-binding proteins to fuse separated catalytic and substrate-binding domains and create new and functional mature proteins. J Med Chem, 1999 Jul 15, 42(14), 2561 - 8 Derivatives of the new ring system indolo{1,2-c}benzo{1,2,3}triazine with potent antitumor and antimicrobial activity; Cirrincione G et al.; Derivatives of the new ring system indolo{1,2-c}benzo{1,2,3}triazine 5 were synthesized by diazotization of substituted 2-(2-aminophenyl)indoles followed by an intramolecular coupling reaction of the diazonium group with the indole nitrogen . To obtain the indolobenzotriazine system it was necessary to protect the 3 position of the indole nucleus to avoid cyclization into the indolo{3,2-c}cinnoline system 4 . Indolobenzotriazines 5a-g were evaluated in vitro for antitumor activity against a panel of leukemia-, lymphoma-, carcinoma-, and neuroblastoma-derived cell lines . Some compounds inhibited the proliferation of T and B cell lines at submicromolar concentrations, whereas their activity against solid tumor cell lines was in the micromolar range . When evaluated for their antifungal potential 5a,d inhibited some of the fungi tested, although at concentrations very close to those inhibiting the proliferation of human cells . On the contrary, all indolobenzotriazines proved fairly potent and selective inhibitors of Streptococcus and Staphylococcus . In particular 5b,c,g were up to 80 times more potent than the reference drug streptomycin and inhibited the growth of the above Gram-positive bacteria at concentrations far lower than those cytotoxic for animal cells. Nihon Kokyuki Gakkai Zasshi, 1999 May, 37(5), 388 - 95 {Comparative study of clinical features of typical and atypical pneumonias}; Watari M et al.; We prospectively analyzed the clinical and laboratory features of 74 patients with community-acquired pneumonia who required hospitalization between May 1996 and October 1997 . Typical pathogens were identified in 47, and atypical pathogens in 27 . The average age was higher in patients affected by typical pathogens (73.9 years), than in patients affected by atypical pathogens (50.9 years) . Univariable analysis found that atypical pneumonias were more frequent in healthy patients than typical pneumonias . Moreover, the presence of relatives with symptoms of airway infection, headache, and earache was more common among the patients with atypical pneumonias, while leukocytosis and elevated C-reactive protein levels were more frequent among patients with typical pneumonias . Typical pathogens accounted for up to 79.6% of the cases of pneumonia with in older patients (aged 60 years or more), whereas atypical pathogens accounted for up to 80% of the cases of pneumonia in younger patients (aged under 60 years) . This difference was statistically significant . Of all 74 patients, 39 (52.7%) were afflicted by severe community-acquired pneumonia, as categorized by American Thoracic Society guidelines . The most common pathogen among these patients was Streptococcus pneumoniae . Legionella was one of the top four . Selection of the initial antimicrobial treatment is an important clinical decision that should be made on the basis of clinical features at admission, age, and severity of the patient's illness. Gastroenterol Hepatol, 1999 Jun-Jul, 22(6), 286 - 9 {Amebic liver abscesses with bacterial superinfection in a nonendemic area}; Ciriza C et al.; A 26-year-old man was admitted to hospital with asthenia, weight loss, right upper quadrant abdominal pain, diarrhea without blood, and fever . Abdominal ultrasonography showed multiple hypoechoic areas in the left hepatic lobe . On abdominal CT, multiple hypodense areas without contrast capture were consistent with hepatic abscesses . Cultures were obtained and the patient was placed empirically on metronidazole, gentamicin and ampicillin, without improvement in the first 72 hours, and a drain was placed in the largest lesion collecting a purulent material . The abscess culture was positive for group C beta-hemolytic Streptococcus . Entamoeba histolytica serology was positive and colon biopsies revealed trophozoites . Multiple left hepatic lobe abscesses secondary to E . histolytica, with bacterial superinfection, is an unusual presentation of amoebic infection, considering that Spain is not an endemic area. Immunopharmacology, 1999 May, 42(1-3), 135 - 49 Genetic disruption of the murine complement C3 promoter region generates deficient mice with extrahepatic expression of C3 mRNA; Circolo A et al.; Genetic deficiencies of the complement protein C3 occur naturally in humans and animal models and have been induced in mice by targeted deletion of the C3 gene . The study of these deficiencies has provided evidence for C3 functions in immune responses . C3 deficient mice were generated by replacing the 5'-flanking region of the C3 gene with the neomycin-resistance (neo) gene . Serum from these mice had no detectable C3 protein or complement activity . Challenge with Streptococcus pneumoniae revealed approximately 2000-fold increase in bacteremia as compared to littermate controls . C3 mRNA was absent in the liver, but it was detected in the lung, kidney, fat tissue, heart and spleen . Metabolic labeling of the lung tissue and peritoneal macrophages showed synthesis of pro-C3, but no post-synthetic intracellular processing of the protein and no secretion of mature C3 . cDNA analysis at the cap site indicated that extrahepatic transcription of the targeted gene was initiated in the neo cassette, close to the C3/neo junction and predicted a primary translation product lacking the leader peptide . The data indicate that these mice provide a good animal model for the study of complete C3 deficiencies and a potential probe for tissue-specific C3 gene regulatory elements. Klin Monatsbl Augenheilkd, 1999 Apr, 214(4), 211 - 6 {Bacterial keratitis in patients with and without contact lens anamnesis}; Frohlich SJ et al.; BACKGROUND: The study shows differences between contact lens wearers and patients without history of contact lenses regarding the spectrum of etiological agents in bacterial keratitis . Based on microbiological analysis, there are given recommendations for an optimal initial antibiotic treatment in both groups . MATERIAL AND METHODS: From 1989 to 1997 smears, scrapings and corneal biopsies were taken from 218 patients with bacterial keratitis . The causing pathogens were isolated on directly inoculated culture media and identified by staining and microscopy . The resistance pattern of a total of 275 germs was analysed for different antibiotics . RESULTS: The most frequently isolated germs were Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus spp., Propionibacterium acnes and Pseudomonas aeruginosa . Whereas sensitive gram-positive germs were predominating in contact lens wearers as well as in non-contact lens wearers, multiresistant gram-negative germs could nearly exclusively be isolated from contact lens wearers . Frequently administered antibiotics like aminoglycosides and quinolones are effective in infections caused by Staphylococcus spp., but increasing resistance could be seen to Streptococcus spp . In this case, erythromycin is very sensitive . Gram-negative germs like Pseudomonas aeruginosa are sensitive to quinolones and some aminoglycosides (e.g . tobramycin) . CONCLUSION: In contact lens wearers, more aggressive germs have to be considered than in non-contact lens wearers . In such cases, frequently administered antibiotics like amino-glycosides are not effective . To cover problematic gram-negative germs we recommend the application of quinolones alternating with erythromycin . The latter one is more effective than quinolones and aminoglycosides in case of Streptococcus spp . co-involvement. Ophthalmology, 1999 Jul, 106(7), 1313 - 8 Emerging fluoroquinolone resistance in bacterial keratitis: a 5-year review; Goldstein MH et al.; OBJECTIVE: To identify resistance patterns to the fluoroquinolones for patients with bacterial keratitis . DESIGN: Retrospective observational case series . PARTICIPANTS: All cases of bacterial keratitis presenting to the Charles T . Campbell Ophthalmic Microbiology Laboratory at the Eye and Ear Institute of Pittsburgh from January 1993 to December 1997 were reviewed . A total of 1053 ocular isolates from 825 cases of bacterial keratitis were identified . MAIN OUTCOME MEASURES: In vitro laboratory susceptibility testing of ocular isolates to ciprofloxacin and ofloxacin was determined by the Kirby-Bauer disk diffusion method and interpreted using the National Committee for Clinical Laboratory Standards serum standards . RESULTS: The number of cases of bacterial keratitis per year decreased from 284 in 1993 to 75 in 1997 . The ratio of gram-positive to gram-negative organisms changed from 81.8%:18.2% in 1993 to 51.4%:48.6% in 1997 (chi-square, 66.00; degrees of freedom, 4; P < 0.000001) . Resistance of Staphylococcus aureus to ciprofloxacin significantly increased annually from 5.8% in 1993 to 35.0% in 1997 (chi-square, 19.80; degrees of freedom, 4; P < 0.0001) and for ofloxacin from 4.7% to 35.0% over the same period (chi-square, 21.32; degrees of freedom, 4; P < 0.001) . Streptococcus species and coagulase-negative Staphylococcus species showed significant resistance to both fluoroquinolones but no change in resistance over the study period . The gram-negative organisms showed good susceptibility to the fluoroquinolones . CONCLUSIONS: This in vitro study shows a significant increased resistance of S . aureus to the fluoroquinolones from 1993 to 1997 . In addition, gaps in fluoroquinolone coverage for Streptococcus and coagulase-negative Staphylococcus species raise concern for the use of monotherapy in treating bacterial keratitis . Contrary to what might be expected, the distribution of gram-positive to gram-negative organisms has shifted, with a decrease in the number of gram-positive organisms identified, while the number of gram-negative isolates has remained stable. J Clin Microbiol, 1999 Aug, 37(8), 2564 - 7 Inhibition enzyme-linked immunosorbent assay for serotyping of group B streptococcal isolates; Arakere G et al.; Group B Streptococcus (GBS) is one of the most common organisms causing neonatal sepsis as well as serious infections in adults . Serotyping the organism is important in studying the epidemiology of the disease as well as deciding a course of treatment . There are several methods available for serotyping . Most of them need high-titered sera and are not quantitative . We are reporting a new inhibition enzyme-linked immunosorbent assay (ELISA) for serotyping which is sensitive and specific compared to the conventional methods but does not need high-titered serotype-specific antisera, as the specificity is controlled by the polysaccharide coating on the ELISA plates . The method can also be quantitative, and we have measured polysaccharide elaborated by different serotype V strains . Thus, the inhibition ELISA method will be useful in serotyping for epidemiological studies, assessing virulence, and performing strain selection for vaccine production. Semin Perinatol, 1999 Jun, 23(3), 250 - 60 Deregulation of cyclooxygenase and nitric oxide synthase gene expression in the inflammatory cascade triggered by experimental group B streptococcal meningitis in the newborn brain and cerebral microvessels; Hauck W et al.; Group B Streptococcus (GBS) is the most common cause of neonatal sepsis and meningitis . Despite antibiotics, GBS in the newborn initiates a cascade of molecular and biological events leading to altered cerebral perfusion, blood-brain barrier disruption, cerebral edema, intracranial hypertension, neurological damage, and even death . Having previously shown that GBS infection impairs cerebral blood flow autoregulation and increases prostaglandin (PG) levels, we examined the regulation of some crucial inflammatory mediators (PGs, nitric oxide (NO), tumor necrosis factor-a) in the brain and cerebral microvessels (MVs) from newborn piglets . Cyclooxygenase (COX), the key enzyme in PG biosynthesis, exists in two isoforms, COX-1 and COX-2 . Both may be directly induced by NO in a model of renal inflammation . Besides its neurotransmitter role, NO is a potent vasorelaxant whose production is catalyzed by at least three distinct nitric oxide synthases (NOS) (bNOS, ecNOS, iNOS) . Western blot analyses showed that the newborn (4 day old) brain expressed lower levels of COX-1 (8-fold), COX-2 (20-fold), bNOS (12-fold), and ecNOS (5-fold) than in the 1 day old . MV showed approximately equal levels of COX-2, lower levels of COX-1 (4-fold), bNOS (5-fold), and higher levels of ecNOS (20-fold) in comparison to 4-day-old cerebral MV . A 4-day-old brain expressed lower levels of bNOS (5-fold), ecNOS (10-fold), and COX-1 (2-fold) than the 6-week-old pig . COX-2 protein was undetected in a 4-day-old pig brain, but present in great excess in MV . Purified MV showed lower ecNOS (14-fold), COX-1 (2-fold), and about equal levels of bNOS and COX-2 in comparison with MV from 6-week-old pigs . Reverse transcription polymerase chain reaction analyses confirmed these results . Treatment with noo-nitro-L-arginine (LNA), a NOS inhibitor, downregulated COX-1 expression in the newborn brain and both COX-1 and COX-2 cerebral MV expression . GBS infection (10(9) colony-forming units, 0.5 mL intracerebroventricular) of sedated newborn piglets induced the expression of tumor necrosis factor-alpha in the cerebrospinal fluid after 2 hours, upregulated bNOS expression in both brain and MVs, upregulated ecNOS in MVs, and downregulated COX-1, COX-2, and ecNOS in the brain . GBS did not trigger the expression of iNOS . Our data suggest that there is a net deficiency of NOS isoforms in the immature brain and microvasculature of the 4-day-old piglet and that the differences in expression lead to the immature control of NO and PG production, rendering newborns particularly susceptible to neurological damage because of the undeveloped nature of their response mechanisms . Moreover, the GBS-induced cascade deregulates the gene expression of interacting inflammatory mediators and may cause a net vasoconstrictor/vasodilator imbalance, leading to cerebral hypertension and edema in the early stages of infection . Pharmacological manipulations of the inflammatory cascade could lead to novel therapeutic approaches for the treatment of GBS meningitis. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 71 - 5 Bactericidal activity of levofloxacin against Streptococcus pneumoniae in an in-vitro model simulating serum pharmacokinetic parameters; Shah PM et al.; The objective of the current study was to evaluate the bactericidal activity of levofloxacin against Streptococcus pneumoniae at concentrations equivalent to those present in serum after a po dosage of 500 mg . Nine S . pneumoniae strains (one penicillin G-resistant, one penicillin G-intermediate resistant, and two penicillin G- and cefotaxime-resistant) were exposed to a levofloxacin concentration of 6 mg/L diluted at a terminal half-life (t1/2) of 8 h . Surviving S . pneumoniae (cfu/mL) were quantified up to 24 h by the membrane filtration method . Levofloxacin was rapidly bactericidal and reduced the quantity of inoculum to below the detection level of 10 cfu/mL within 2.5-5.15 h, irrespective of susceptibility to penicillin G or cefotaxime . No viable S . pneumoniae could be detected at the end of the observation period (24 h) . All strains except one (strain 17134) had an MIC < 1.0 mg/L, and the minimum bactericidal concentrations (MBCs) were, at the most, one dilution higher than the respective MICs . The inoculum was high, ranging from 2.9 x 10(5) to 7.5 x 10(6) cfu/mL . The time required to achieve 99% death ranged from 0.9 to 3.1 h, and was longest for strain 17134 which had an MIC of 1.0 mg/L and an MBC of 2.0 mg/L . A 99.9% reduction in inoculum was achieved within 1.5-4.15 h . At a serum concentration achievable after a single po dosage of 500 mg, levofloxacin showed rapid and complete bactericidal activity against the S . pneumoniae strains tested. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 61 - 5 Serum bactericidal activity of levofloxacin against Streptococcus pneumoniae; Shah PM; The objective of this study was to determine the serum bactericidal activity (SBA) of levofloxacin against Streptococcus pneumoniae strains with various degrees of susceptibility to penicillin and cefotaxime . Serum samples of volunteers (n = 12) who had received levofloxacin 500 mg as a single po dose were provided in blinded fashion . SBA was determined, using the microdilution method, in Todd-Hewitt broth supplemented with lysed horse blood inoculated with an overnight culture diluted to yield a final concentration of approximately 10(5) cfu/mL . The serum bactericidal titre was defined as the highest dilution of serum showing no growth (> 99.9% reduction of inoculum) . The duration of SBA ranged from 0.75 to 6.3 h (mean 3.85 h), and was independent of the susceptibility of the strains to penicillin and cefotaxime . In conclusion, a single po dose of 500 mg levofloxacin achieved serum concentrations which were bactericidal against penicillin-resistant S . pneumoniae for a mean period of 3.85 h. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 55 - 9 Relative potential for selection of fluoroquinolone-resistant Streptococcus pneumoniae strains by levofloxacin: comparison with ciprofloxacin, sparfloxacin and ofloxacin; Drugeon HB et al.; The aim of this study was to evaluate the relative potential of levofloxacin to select for resistance in Streptococcus pneumoniae in comparison with ciprofloxacin, sparfloxacin and ofloxacin . Two S . pneumoniae strains were studied; HBD 153 (parental strain, serotype 3) and HBD 964 (selected from the parental strain in an experimental mouse peritonitis infection model) . MICs for the two strains were, respectively: 2 and 2 mg/L for ciprofloxacin; 2 and 4 mg/L for ofloxacin; 0.5 and 1 mg/L for sparfloxacin; 2 and 2 mg/L for levofloxacin . In-vitro, with 4 x MIC as the selection concentration, no mutant was obtained with strain HBD 153 (mutation frequency < 10(-8) . With HBD 964, the mutation frequency was 9 x 10(-7) for ofloxacin, 10(-7) for ciprofloxacin, 4 x 10(-5) for sparfloxacin and < 10(-8) for levofloxacin . In an immunosuppressed mouse peritonitis model (20 mice per dose), the S . pneumoniae strains were studied with sc doses of ciprofloxacin, sparfloxacin and ofloxacin at 50 mg/kg od, and with sc levofloxacin at a dose of 10 and 50 mg/kg od, or 10 and 50 mg/kg bid . The MICs for strains isolated after antibiotic treatment and the mutation frequencies at 4 x MIC were determined . Against HBD 153, sparfloxacin was the most active treatment, followed by levofloxacin 10 mg/kg and 50 mg/kg bid, but strains identical to HBD 964 (showing a resistant variant at 4 x MIC) were selected by sparfloxacin . Against HBD 964, levofloxacin (10 mg/kg and 50 mg/kg) was the most active drug . Highly resistant mutants were selected by ofloxacin and ciprofloxacin, but not by sparfloxacin and levofloxacin . In conclusion, the relative potential of levofloxacin to select for fluoroquinolone-resistant S . pneumoniae is lower than that of ciprofloxacin, ofloxacin and sparfloxacin both in vitro and in vivo. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 15 - 9 In-vitro activity of levofloxacin against Streptococcus pneumoniae with various levels of penicillin resistance; Thomson KS et al.; This in-vitro study was designed to compare the activity of levofloxacin with that of ciprofloxacin, ofloxacin, erythromycin, penicillin, amoxycillin, loracarbef, cefaclor, cefpodoxime, ceftriaxone, trimethoprim-sulphamethoxazole, clindamycin and vancomycin against a collection of 202 Streptococcus pneumoniae isolates (56% susceptible to penicillin, 34% intermediate, 10% resistant) . The isolates (60% nasopharyngeal, 40% middle ear) were obtained from otherwise healthy children at child care centres in urban and rural Nebraska, and at a paediatric clinic in rural Kentucky . MICs were determined by NCCLS agar dilution methodology using an inoculum of 10(4) cfu/spot . Using NCCLS breakpoints, the percentage of penicillin-intermediate and -resistant strains susceptible to the evaluable agents were, respectively, as follows: levofloxacin (99%, 100%), ofloxacin (87%, 100%), erythromycin (52%, 65%), ceftriaxone (93%, 25%), trimethoprim-sulphamethoxazole (7%, 0%), clindamycin (93%, 100%) and vancomycin (100%, 100%) . Without NCCLS interpretive criteria, no conclusions could be made concerning the susceptibility of penicillin-intermediate and -resistant strains to the other study drugs . All beta-lactam antibiotics, erythromycin and trimethoprim-sulphamethoxazole were less active against penicillin-resistant strains, indicating that these may be suboptimal agents for empirical therapy for suspected S . pneumoniae infections in these patient populations . However, levofloxacin, ofloxacin, clindamycin and vancomycin were equally active against penicillin-susceptible and -resistant strains . These data suggest that the efficacy of levofloxacin should be examined in both adult and paediatric S . pneumoniae infections involving body sites where levofloxacin concentrations > 2 mg/L can be achieved safely. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 9 - 14 In-vitro bacteriostatic activity of levofloxacin and three other fluoroquinolones against penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae; Fremaux A et al.; The purpose of this study was to investigate the in-vitro bacteriostatic activity of levofloxacin in comparison with that of ofloxacin, sparfloxacin and ciprofloxacin against 205 strains of Streptococcus pneumoniae (101 penicillin-susceptible, 51 penicillin-intermediate and 53 penicillin-resistant) . The isolates were provided between September 1996 and October 1996 by French hospitals participating in the National Co-operative Survey of Pneumococcal Infections . The determination of MICs (mg/L) was made by the agar dilution method . The MIC50 and MIC90 values of the four fluoroquinolones for the three classes of S . pneumoniae (penicillin-susceptible, penicillin-intermediate and penicillin-resistant) were not significantly different . In contrast, the differences in in-vitro activity observed among the four fluoroquinolones against the 205 strains allowed them to be separated into three groups: sparfloxacin (MIC50/90 0.25 mg/L); ciprofloxacin and levofloxacin (MIC50 0.5 and 1 mg/L respectively, MIC90 1 mg/L); and ofloxacin (MIC50 1 mg/L, MIC90 2 mg/L) . A total of 204 of the strains had a levofloxacin MIC between 0.25 mg/L and 1 mg/L, and only one of the 205 strains was highly resistant (MIC 16 mg/L) . Whatever the level of susceptibility to penicillin, the relative bacteriostatic activity was, in descending order of activity, sparfloxacin, levofloxacin/ciprofloxacin and ofloxacin . These results suggest levofloxacin has potential for the treatment of pneumococcal infections. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 5 - 8 A comparative study of the in-vitro activity of levofloxacin against Streptococcus pneumoniae; Reinert RR et al.; In this study, the in-vitro activity of levofloxacin against Streptococcus pneumoniae was compared with the activities of a range of other antibiotics . In total, 320 penicillin-susceptible and 30 penicillin-intermediate clinical isolates of S . pneumoniae were collected in Germany between 1992 and 1994 from patients with bacteraemic pneumonia . MICs were determined using the agar dilution methodology recommended by the NCCLS and the results with levofloxacin compared with those with ofloxacin, D-ofloxacin, ciprofloxacin, amoxycillin, cefpodoxime, cefixime, cefuroxime, faropenem, erythromycin and tetracycline . Levofloxacin (MIC50 1 mg/L) was approximately twice as active against the isolates as ofloxacin (MIC50 2 mg/L) . D-ofloxacin (MIC90 32 mg/L) showed no activity, while beta-lactam antibiotics showed elevated MIC90 values against penicillin-intermediate strains (amoxycillin, 1 mg/L; cefpodoxime, 2 mg/L; cefixime, 32 mg/L; cefuroxime, 8 mg/L) in comparison with the MIC90 obtained with penicillin-susceptible strains (amoxycillin, 0.015 mg/L; cefpodoxime, 0.03 mg/L; cefixime, 0.5 mg/L; cefuroxime, 0.03 mg/L) . Faropenem showed good activity against pneumococcal isolates (penicillin-susceptible strains, MIC90 0.016 mg/L; penicillin-intermediate strains, MIC90 0.25 mg/L) . Erythromycin (MIC90 8 mg/L) and tetracycline (MIC90 64 mg/L) were also less active against penicillin-intermediate pneumococcal isolates . In conclusion, levofloxacin and faropenem may be useful in the treatment of pneumococcal infections caused by organisms with decreased susceptibility to penicillin. J Antimicrob Chemother, 1999 Jun, 43 Suppl C, 1 - 3 The susceptibility of Streptococcus pneumoniae to levofloxacin and other antibiotics; Keller N et al.; The aim of this study was to determine the in-vitro susceptibility of Streptococcus pneumoniae to levofloxacin, ciprofloxacin, sparfloxacin, ofloxacin, amoxycillin, cefepime and cefuroxime . In total, 105 isolates (clinical blood and sputum isolates from 1995-96) were selected from the collection of the Department of Clinical Microbiology of the Chaim Sheba Medical Centre . MICs were determined with an agar dilution method according to NCCLS guidelines . The MIC ranges with the test antibiotics against the pneumococcal isolates were: 0.5 mg/L, levofloxacin; 0.12-0.5 mg/L, sparfloxacin; 0.5-2 mg/L, ciprofloxacin; 0.5-1 mg/L, ofloxacin; < 0.03-2 mg/L, amoxycillin; < 0.03-2 mg/L, cefepime; < 0.03-2 mg/L, cefuroxime . The results indicated that although all isolates were susceptible to all the fluoroquinolone agents except ciprofloxacin, some isolates had reduced susceptibility to amoxycillin, cefepime and cefuroxime . We conclude that all S . pneumoniae isolates tested were susceptible to levofloxacin, ofloxacin and sparfloxacin . However, about 85% were susceptible to amoxycillin and the other beta-lactam antibiotics tested. J Antimicrob Chemother, 1999 Jun, 43(6), 833 - 5 Comparative in-vitro activity of moxifloxacin, penicillin, ceftriaxone and ciprofloxacin against pneumococci isolated from meningitis; Tarasi A et al.; Minimum inhibitory concentrations of penicillin, ceftriaxone, ciprofloxacin, and moxifloxacin (BAY 12-8039), a new 8-methoxyquinolone, were determined for 60 cerebrospinal fluid isolates of Streptococcus pneumoniae collected during January 1997-April 1998 at Italian medical centres . Three reference isolates with predetermined MIC values (two penicillin- and multidrug-resistant isolates, one uniformly susceptible to all antibiotics) were also tested with the same antibiotics . The MIC90 of penicillin was < or = 0.03 mg/L (range < or = 0.03-2 mg/L), of ceftriaxone 0.06 mg/L (range < or = 0.03-0.5 mg/L), of ciprofloxacin 2 mg/L (range 0.5-8 mg/L) and of moxifloxacin 0.06 mg/L (range 0.03-0.12 mg/L) . Moxifloxacin was effective against all the penicillin-resistant isolates tested, with an MIC of 0.06 mg/L . Moxifloxacin was 32-fold more active than ciprofloxacin and was not affected by penicillin and cephalosporin resistance . These results indicate that moxifloxacin could be useful for the treatment of both penicillin-sensitive and -resistant S . pneumoniae meningitis. J Antimicrob Chemother, 1999 Jun, 43(6), 811 - 6 Pharmacodynamics of trovafloxacin in a mouse model of cephalosporin-resistant Streptococcus pneumoniae pneumonia; Ng W et al.; Trovafloxacin is a potentially useful agent for treatment of infections caused by cephalosporin-resistant Streptococcus pneumoniae . We studied the effectiveness of trovafloxacin therapy and examined the correlation between pharmacodynamic indices in serum and lung, and bacterial killing . Immunocompetent Balb/c mice were infected by intranasal inoculation of a cephalosporin-resistant S . pneumoniae isolate (MIC of ceftriaxone and trovafloxacin 2 and 0.06 mg/L, respectively) . Trovafloxacin 10-30 mg/kg/day in one or three divided doses was started 15 h after infection . Serum and lung drug concentrations were measured at multiple time points for 24 h . Serum concentrations peaked at 30-60 min and lung concentrations approximately 30 min later . The serum T1/2 was approximately 9 h and lung T1/2 varied from 5 to 9 h . Lung AUC and Cmax values were 2-3 times greater than those in serum . At the start of therapy lung bacterial concentrations were 8.4 +/- 0.3 log10 cfu/mL and 24 h later had decreased by 3.5 +/- 0.2, 4.0 +/- 0.2, 0.8 +/- 0.3 and 1.0 +/- 1.2 log10 cfu/mL with 30 mg/kg x 1, 10 mg/kg x 3, 10 mg/kg x 1 and 3.3 mg/kg x 3 regimens, respectively . Although the larger dosages were more effective (P < 0.001) the differences between divided and single dosage regimens were not significant . Trovafloxacin serum AUC/MIC ratio correlated best with bacterial killing in the lungs over 24 h . Trovafloxacin is likely to be useful in the treatment of cephalosporin-resistant S . pneumoniae pneumonia. J Antimicrob Chemother, 1999 Jun, 43(6), 783 - 92 Prevalence of resistance to MLS antibiotics in 20 European university hospitals participating in the European SENTRY surveillance programme . Sentry Participants Group; Schmitz FJ et al.; Macrolide, lincosamide and streptogramin (MLS) antibiotics are chemically distinct inhibitors of bacterial protein synthesis . Resistance to MLS antibiotics may be constitutive or inducible . The purpose of this study is to update our understanding of the prevalence of different forms of MLS resistance in Europe . The analysis of 3653 clinical pneumococcal, staphylococcal and enterococcal isolates exhibited an average percentage of 21.3% and 6.2% intermediate and high-level penicillin-resistant Streptococcus pneumoniae, 21.8% methicillin-resistant Staphylococcus aureus and 11% vancomycin-resistant Enterococcus faecium . Geographical differences in erythromycin and clindamycin resistance in isolates of S . pneumoniae and S . aureus strongly reflect geographical variations in susceptibility to penicillin and methicillin, respectively . A very narrow range of MICs was obtained with quinupristin/dalfopristin, with no S . pneumoniae, S . aureus and E . faecium isolate having an MIC of > 4 mg/L, indicating a possible role of quinupristin/dalfopristin in the treatment of infections by multi-resistant Gram-positive bacteria. J Antimicrob Chemother, 1999 Jun, 43(6), 777 - 82 Differences between the activity of penicillin, amoxycillin, and co-amoxyclav against 5,252 Streptococcus pneumoniae isolates tested in the Alexander Project 1992-1996; Butler DL et al.; A number of published studies have shown that the MICs of amoxycillin and/or co-amoxyclav are lower than those of ampicillin and/or penicillin for Streptococcus pneumoniae . Other published studies have concluded that the activities of amoxycillin and co-amoxyclav are comparable with that of penicillin for S . pneumoniae . A collection of 5252 S . pneumoniae isolates obtained during a 5 year period (1992-1996) was analysed to determine differences between the MICs of penicillin, amoxycillin and co-amoxyclav . Among the isolates analysed, 3788 (72%) were penicillin-susceptible, 615 (12%) were penicillin-intermediate and 849 (16%) were penicillin-resistant . Differences between the agents were assessed by examination of MIC distribution functions and simultaneous 95% CIs . In addition, penicillin-intermediate and -resistant isolates were analysed to determine the number and percentage of isolates which had an amoxycillin and co-amoxyclav MIC less than, equal to, or greater than the penicillin MIC . Results showed that the amoxycillin and co-amoxyclav MIC90s were one two-fold dilution lower than those of penicillin for all isolates collected between 1992-1993 and 1994-1996 . Simultaneous 95% CIs showed that the mean differences between MICs of amoxycillin and penicillin, and between MICs of co-amoxyclav and penicillin, were less