An Pediatr (Barc), 2003 Dec, 59(6), 565 - 70 {Cost benefit analysis of body surface cultures at a neonatal unit}; Marugan Isabel V et al.; INTRODUCTION: Infection continues to be a cause of concern in neonatal units . The high cost of the body surface cultures used to study infection and their limited utility is controversial . MATERIAL AND METHODS: The medical records of newborns admitted for suspected infection in 1999 were retrospectively reviewed . Request criteria, cost, and the clinical utility of body surface cultures were analyzed . RESULTS: In 1999, body surface cultures were requested in 204 newborns admitted to hospital (70 % of all admitted newborns) . Of these, six were diagnosed with bacteremia (6.23/1000; 95 % CI: 5.9-6.5) . The most frequently isolated microorganisms were Escherichia coli and Streptococcus agalactiae . The total cost of body surface cultures was 6,510.95 euros (1,083,331 pesetas) . In 25 % of cases the results of cultures influenced clinical decision making . The cost necessary to obtain clinical repercussion in a patient was 191.50 euros (31,863 pesetas) . Requesting two body surface cultures only (otic and umbilical) halved the cost without diminishing their clinical utility . CONCLUSIONS: A considerable percentage of newborns admitted to hospital present suspected infection requiring microbiological studies . Although the cost of body surface cultures is high, the results of these cultures influence diagnostic and therapeutic decisions in a quarter of patients . We do not believe that eliminating the use of these cultures would be beneficial . However, their cost can be reduced by carefully selecting request criteria and by limiting cultures to two samples (otic and umbilical). Eur J Immunol, 2003 Dec, 33(12), 3322 - 30 Critical role of Valpha14+ natural killer T cells in the innate phase of host protection against Streptococcus pneumoniae infection; Kawakami K et al.; The present study was designed to elucidate the role of Valpha14(+) NKT cells in the host defense against pulmonary infection with Streptococcus pneumoniae using Jalpha281 gene-disrupted mice (Jalpha281KO mice) that lacked this lymphocyte subset . In these mice, pneumococcal infection was severely exacerbated, as shown by the shorter survival time and marked increase of live bacteria in the lung compared to wild-type (WT) mice . The proportion of Valpha14(+) NKT cells, detected by an alpha-galactosylceramide (alpha-GalCer)-loaded CD1d tetramer, increased in thelung after S . pneumoniae infection . This increase was significantly reduced in mice with a genetic disruption of monocyte chemotactic protein (MCP)-1, which was produced in the early phaseof infection in WT mice . In the lungs of Jalpha281KO mice, the number of neutrophils was significantly lower at 12 h than that in WT mice . In support of this finding, macrophage inflammatory protein (MIP)-2 and TNF-alpha synthesis in infected lungs was significantly reduced at 3 h and at both 3 and 6 h, respectively, in Jalpha281KO mice, compared to WT mice . In addition, treatment of mice with alpha-GalCer significantly improved the outcome of this infection . Our results demonstrated MCP-1-dependent recruitment of Valpha14(+) NKT cells and their critical role in early host protection against S . pneumoniae by promoting the trafficking of neutrophils to the site of infection. Alcohol Clin Exp Res, 2003 Nov, 27(11), 1838 - 45 Alcohol-induced suppression of lung chemokine production and the host defense response to Streptococcus pneumoniae; Boe DM et al.; BACKGROUND: Acute alcohol intoxication impairs neutrophil migration in response to intrapulmonary infection with Streptococcus pneumoniae, the most common bacterial cause of pneumonia . Many of the same host defense functions that are impaired in the alcohol-intoxicated host are mechanistically associated with chemokines, a group of proinflammatory molecules that enhance neutrophil adhesion and direct neutrophil migration to sites of inflammation . The purpose of this study was to determine whether alcohol-induced chemokine suppression is responsible for impaired neutrophil recruitment into the lung during infection of the alcohol-intoxicated host . METHODS: S . pneumoniae was administered (107 colony-forming units) intratracheally 30 min after intraperitoneal injection of 20% alcohol (5.5 g/kg) or saline . Four hours after bacterial challenge, bronchoalveolar lavage fluid (BALF) was collected, and the ability of BALF to induce neutrophil chemotaxis and adhesion molecule expression was measured by using chemotactic and flow cytometric assays . In another experiment, intratracheal challenge was performed by using recombinant macrophage inflammatory protein-2 (MIP-2), and BALF neutrophils were measured . RESULTS: BALF MIP-2 and cytokine-induced neutrophil chemoattractant were decreased by alcohol, and BALF from alcohol-intoxicated animals had decreased chemotactic activity for neutrophils, as well as a decreased ability to up-regulate neutrophil adhesion molecule expression, compared with controls . This decreased chemotactic activity was significantly increased in the alcohol group by repletion of chemokines to control levels . Alcohol also suppressed neutrophil recruitment after intrapulmonary challenge with MIP-2, suggesting that mechanisms other than chemokine suppression contribute to the alcohol-induced effect . CONCLUSIONS: At least two mechanisms, suppressed chemokine production and impaired neutrophil adhesion molecule expression, likely work in concert in the alcohol-intoxicated host to impair neutrophil adhesion and migration into the lung during pneumococcal infection . These alterations in neutrophil function likely increase the susceptibility of alcohol-consuming hosts to pneumonia. Microb Drug Resist, 2003, 9 Suppl 1, S61 - 8 Inferring the potential success of pneumococcal vaccination in Italy: serotypes and antibiotic resistance of Streptococcus pneumoniae isolates from invasive diseases; Pantosti A et al.; To evaluate the potential impact of antipneumococcal vaccination in Italy, Streptococcus pneumoniae isolates from invasive disease were collected from 65 laboratories in the years 1997-2000 . Of the 503 isolates examined, 15% were from children <5 years and 34% from adults > or = 65 years . The most frequent serogroups were, in ranking order, 14, 19, 6, and 23 . Overall, 93.8% of the isolates belonged to serogroups enclosed in the 23-valent polysaccharide vaccine . Among children isolates, serotypes 14, 6B, and 23F comprised 60% of the isolates; overall, 72% of the isolates belonged to serotypes included in the heptavalent conjugate vaccine . Penicillin nonsusceptible isolates (10%) belonged to a limited number of serogroups, being more common in serogroups 19 and 9 and in the nonvaccine serogroups 24 and 35 . Erythromycin-resistant isolates (29%) belonged to several serogroups, more frequently to serogroups 14, 6, and 19 . Both vaccines are potentially able to prevent the majority of resistant infections in the respective age groups in Italy. Microb Drug Resist, 2003, 9 Suppl 1, S53 - 9 Experimental study of clinafloxacin alone and in combination in the treatment of ciprofloxacin-susceptible and -resistant pneumococcal meningitis; Domenech A et al.; The increasing incidence of ciprofloxacin resistance in Streptococcus pneumoniae may limit the efficacy of the new quinolones in difficult-to-treat infections such as meningitis . The aim of the present study was to determine the efficacy of clinafloxacin alone and in combination with teicoplanin and rifampin in the therapy of ciprofloxacin-susceptible and ciprofloxacin-resistant pneumococcal meningitis in rabbits . When used against a penicillin-resistant ciprofloxacin-susceptible strain (Clinafloxacin MIC 0.12 microg/ml), clinafloxacin at a dose of 20 mg/kg per day b.i.d . decreased bacterial concentration by -5.10 log cfu/ml at 24 hr . Combinations did not improve activity . The same clinafloxacin schedule against a penicillin- and ciprofloxacin-resistant strain (Clinafloxacin MIC 0.5 microg/ml) was totally ineffective . Our data suggest that a moderate decrease in quinolone susceptibility, as indicated by the detection of any degree of ciprofloxacin resistance, may render these antibiotics unsuitable for the management of pneumococcal meningitis. Microb Drug Resist, 2003, 9 Suppl 1, S35 - 8 Phenotypic and genetic characterization of resistance against macrolides and lincosamides in Streptococcus gallolyticus strains isolated from pigeons and humans; Kimpe A et al.; The macrolide and lincosamide (ML) resistance phenotype of 65 pigeon and 30 human Streptococcus gallolyticus strains was determined by the disk diffusion method . Constitutive resistance against the tested antibiotics was seen in 13 human and 28 pigeon strains . Simultaneous screening for the presence of erm(B) and mef(A) genes using PCR revealed that the erm(B) gene was present in 40 out of these 41 phenotypically resistant S . gallolyticus strains while the mef(A) gene was detected in only one resistant and one susceptible human-derived strain . The erm(B) genes of 10 human and 10 pigeon S . gallolyticus strains were sequenced and compared . Four human and seven pigeon strains possessed exactly the same sequence for the erm(B) gene . The sequence of the erm(B) gene of the remaining strains differed in one to five nucleotides . These findings could indicate a possible exchange of resistance genes between human and pigeon strains. Lett Appl Microbiol, 2003, 37(5), 370 - 3 Detection of Streptococcus anginosus from saliva by real-time polymerase chain reaction; Kumagai K et al.; AIMS: The purpose of the present investigation was to assess the salivary levels of Streptococcus anginosus in periodontitis patients . METHODS AND RESULTS: The salivary levels of Strep . anginosus were assessed using real-time polymerase chain reaction (PCR) . Streptococcus anginosus was detected in 28 of 37 (75.6%) of periodontitis patients and in three of the 20 (15%) healthy subjects . The mean values for bleeding on probing and probing depth in positive patients were statistically higher than those in negative patients . A significant decrease in Strep . anginosus levels was observed after periodontal treatment . CONCLUSIONS: Although the levels of Strep . anginosus are extremely low, they may reflect the status of periodontal health . SIGNIFICANCE AND IMPACT OF THE STUDY: Real-time PCR is a useful method for obtaining the relative quantities of Strep . anginosus from saliva samples and for monitoring the effect of therapy. Paediatr Drugs, 2003, 5 Suppl 1, 35 - 46 Acute bacterial skin infections in pediatric medicine: current issues in presentation and treatment; Hedrick J; Bacterial skin and skin structure infections commonly encountered in children include impetigo, folliculitis, furunculosis, carbuncles, wound infections, abscesses, cellulitis, erysipelas, scarlet fever, acute paronychia, and staphylococcal scalded skin syndrome . If diagnosed early and treated appropriately, these infections are almost always curable, but some have the potential to cause serious complications such as septicemia, nephritis, carditis and arthritis if diagnosis is delayed and/or treatment is inadequate . During the initial evaluation, it is important to determine whether the infection is superficial or deep, and whether it is localized or spreading . Prompt treatment is essential if the infection appears to be spreading, as the sequelae can be life threatening . Once the proper diagnosis is made, the next important step is selecting the most appropriate therapy . In children presenting with mild or moderately severe bacterial skin and skin structure infections and not requiring inpatient management or urgent operative debridement, prompt provision of oral antimicrobial therapy avoids the risk of worsening infection or hospitalization . Empiric antimicrobial therapy should be directed at the most likely pathogens, (e.g . Staphylococcus aureus or Streptococcus pyogenes), although some infections (e.g . subcutaneous abscesses and cellulitis following animal or human bites) may have a polymicrobial origin . In choosing the appropriate antimicrobial therapy, one must take into account the resistance profile of the target pathogen, the agent's antibacterial profile and intrinsic activity against the target pathogen, and its pharmacokinetic properties (including absorption, elimination, and extent of tissue penetration) . Other factors to consider include tolerability of the drug, convenience of the dosing regimen, and acceptability and palatability of the oral formulation administered . Any treatment plan for bacterial skin and skin structure infections should aim to minimize the emergence of resistant organisms so that the risk of their dissemination to others in the community is reduced . Oral antimicrobial agents currently available that may be considered include: beta-lactamase-stable penicillins (e.g . cloxacillin, dicloxacillin, and amoxicillin-clavulanate potassium), the macrolides (e.g . erythromycin, clarithromycin, and azithromycin), and the cephalosporins . Cephalosporins are now the most commonly used class, particularly because of increasing resistance among strains of S . pyogenes to erythromycin (and by implication, the other macrolides) . The second- and third-generation cephalosporins have many advantages, with their extended spectra of antimicrobial activity, favorable pharmacokinetic and tolerability profiles, and convenient dosage schedules . The third-generation agent, cefdinir, has good activity against a broad range of likely pathogens, including staphylococci, a twice-daily administration schedule, a favorable efficacy and tolerability profile, is well accepted by young children when administered as an oral suspension, and may be an attractive alternative in the pediatric setting. Paediatr Drugs, 2003, 5 Suppl 1, 13 - 23 Acute streptococcal pharyngitis in pediatric medicine: current issues in diagnosis and management; Shulman ST; Group A beta-hemolytic streptococcus (GABHS) is the most common bacterial cause of acute pharyngitis . Although children infected with GABHS will recover clinically without antibiotics, treatment is recommended in order to prevent acute rheumatic fever and probably suppurative complications, hasten resolution of clinical signs and symptoms, and prevent transmission to close contacts . Streptococcal pharyngitis usually cannot be reliably distinguished from other etiologies on the basis of epidemiologic or physical findings, and therefore a throat culture or a rapid antigen detection test is generally necessary to confirm the diagnosis . All isolates of GABHS are sensitive to penicillins and cephalosporins, whereas resistance to macrolides has been identified in some geographic regions . The recommended first-line therapy for streptococcal pharyngitis is a 10-day course of penicillin V, usually given 2 or 3 times per day . A number of alternatives to penicillin V are available, including other penicillins, macrolides, and cephalosporins . As a class, the cephalosporins are noteworthy because they may provide somewhat higher bacteriologic eradication rates than penicillin V . Many cephalosporins can be administered twice daily, but they also must be given for 10 days . Two third-generation cephalosporins, cefdinir and cefpodoxime proxetil, are approved for use in a more convenient 5-day dosing schedule, thus possibly increasing the likelihood of adherence to the full course of therapy . Palatability is also an important consideration when prescribing antibiotics to children . In a series of studies, children preferred the pleasant strawberry-cream taste of cefdinir to that of amoxicillin/clavulanate, cefprozil, and azithromycin . Cefdinir may offer an alternative to penicillin V for children with streptococcal pharyngitis, particularly when compliance is a clinical concern. Presse Med, 2003 Sep 6, 32(28 Suppl), S24 - 6 {Following the serotypes of pneumococcal strains isolated in asplenic subjects}; Varon E; EPIDEMIOLOGY: There have been relatively few studies concerning the epidemiology of Streptococcus pneumoniae infections in asplenic subjects, and even fewer concerning pneumococcal carriage . A small number of studies provide data on serotype frequency, or simply on serogroups . The frequency of pneumococcal carriers among sickle-cell anemia children varies from 6% to 27% depending on the series and the type of sampling used (rhinopharyngeal or oropharyngeal) . IMPORTANCE OF SURVEILLANCE: It is particularly important to carefully follow the evolution of the serotypes and sensitivities of pneumococci isolated from carriers in order to assess the selective pressure of the antibiotic prophylaxis and to determine the impact of anti-pneumococcal vaccination in this population, as in the general population. Infect Dis Obstet Gynecol, 2003, 11(2), 117 - 22 Second-trimester pregnancy loss at an urban hospital; Heller DS et al.; OBJECTIVES: Second-trimester spontaneous pregnancy losses are less common than first-trimester losses, and are often associated with ascending infection and/or acute chorioamnionitis . A Medline search revealed only two large studies published in the recent literature, reporting incidences of chorioamnionitis of 39.3% and 58.2%, respectively . These studies did not address the use of histopathology for the identification of organisms . Since ascending infection is likely to be a significant cause of second-trimester loss in the inner-city population at the University Hospital in Newark, New Jersey, we sought to evaluate the usefulness of stains for microorganisms, which are rarely utilized on these specimens . METHODS: Retrospective review of the medical records and pathologic material for cases of spontaneous abortions seen at the University Hospital in Newark between January 1999 and March 2001 was undertaken . Stains for microorganisms were performed on archival placental tissue for cases with histologic acute chorioamnionitis . RESULTS: A total of 67 cases were available for review, of which 38 cases (56.7%) showed histologic acute chorioamnionitis, similar to the rates in one previous study, but significantly higher than those in the other (p = 0.01) . Of 25 cases with histological chorioamnionitis for which appropriate fetal material was available, 13 cases (52%) showed polymorphonuclear leukocytes (PMNs) in the fetal lungs, one case (4%) showed PMNs in the fetal stomach, and seven cases (28%) showed PMNs in both the lung and the stomach . Of the 38 cases with chorioamnionitis, Gram stains showed Gram-positive cocci in six cases, two of which were culture positive for group B streptococcus . Warthin-Starry stains showed filamentous organisms consistent with Fusobacterium sp . in the placenta in three cases . CONCLUSIONS: Acute chorioamnionitis is associated with second-trimester pregnancy loss at this inner-city hospital, and may be related to the high incidence of risk factors in this population . A small proportion of cases can be further characterized by the inclusion of Gram and Warthin-Starry stains in the evaluation . Selection of cases with histologic acute chorioamnionitis for further study with special stains may provide additional information on the causative organism. Int J Clin Pract, 2003 Oct, 57(8), 735 - 6 A case of septicaemia, meningitis and pneumonia caused by Streptococcus bovis type II; Namiduru M et al.; We present a case of septicaemia, pneumonia and meningitis due to Streptococcus bovis type-II in a patient who had undergone a total hip prosthesis under general anaesthesia three weeks earlier . This organism is an uncommon human pathogen that sometimes causes bacteraemia and endocarditis and is usually connected with colon pathology and dental procedures . In the reported case, there were no risk factors for S . bovis infection except for the hip operation . S . bovis type II sensitive to penicillin was isolated from all blood and pleural fluid cultures . The patient recovered and was discharged from hospital two weeks after presentation. Nat Biotechnol, 2003 Dec, 21(12), 1480 - 5 Epub 2003 Nov 16. An engineered multidomain bactericidal peptide as a model for targeted antibiotics against specific bacteria; Qiu XQ et al.; We constructed a peptide consisting of a staphylococcal AgrD1 pheromone fused to the channel-forming domain of colicin Ia and named it pheromonicin . This fusion peptide had bactericidal effects against methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA, respectively), but not against Staphylococcus epidermidis or Streptococcus pneumoniae . Growth rates, vital staining and colony forming unit (CFU) counts showed that pheromonicin did not merely suppress growth but killed S . aureus cells . The specificity of pheromonicin was shown by the absence of bactericidal effects against an accessory gene regulator (agr) locus knockout of S . aureus, and a dose-dependent inhibition of the bactericidal effects of pheromonicin by competition with corresponding free AgrD pheromone . In vivo, all pheromonicin-treated mice survived administration of MRSA that was lethal to controls . No toxicity was detectable in human liver or renal cells in culture, or in livers, kidneys or spleens of pheromonicin-treated mice . The results suggest that these types of chimeric peptides may be of value as antibiotics against specific bacterial infections. J Infect Dis, 2003 Nov 15, 188(10), 1587 - 92 Epub 2003 Nov 10. Array of M protein gene subtypes in 1064 recent invasive group A streptococcus isolates recovered from the active bacterial core surveillance; Li Z et al.; Using sequence analysis to detect variation within the hypervariable M protein N terminus, we found 41 emm types encompassing 81 subtypes, among 1064 consecutive invasive group A streptococcus isolates from a recent multistate, population-based surveillance . Seventeen of the 30 emm types represented by multiple isolates displayed multiple subtypes . Most subtypes differed from reference strain emm sequences as a result of single base substitutions or other alterations likely to be stably inherited . The Centers for Disease Control and Prevention database (available at: currently contains 225 distinct emm types encompassing 450 subtypes . Although this subtyping scheme increases specificity, limited variation within individual types favors introduction of M protein type-specific vaccines. Mol Microbiol, 2003 Nov, 50(4), 1103 - 10 From nose to lung: the regulation behind Streptococcus pneumoniae virulence factors; Hava DL et al.; Streptococcus pneumoniae probably possesses a redundant set of factors required for colonization of the nasopharynx and invasive disease, because of its strict relationship with its human host and relatively small genome size (approximately 2.1 Mb) . Nevertheless, transcriptional regulation of genes encoding factors required for in vivo growth is predicted to be important on two fronts: in the transition from carriage to invasive disease and within different microniches of the nasopharynx . The importance of both serotype-specific and host tissue-specific virulence factors during infection and disease has been highlighted by the recent identification of novel virulence factors in this organism coupled with the release of complete genome sequences from two strains . These studies add to the foundation of knowledge of classical S . pneumoniae virulence factors such as polysaccharide capsule and pneumolysin, which have well-documented roles in pathogenesis. Oral Microbiol Immunol, 2003 Dec, 18(6), 371 - 8 A rapid procedure to ascertain the antimicrobial efficacy of oral care formulations; Sreenivasan PK et al.; A rapid method examining the antimicrobial efficacy of oral care formulations with alamar blue, an oxidation-reduction dye with fluorescent end-points, is described . Significant correlations between increasing viable plate counts of the oral bacteria Actinomyces viscosus, Streptococcus sanguis, Streptococcus mutans and Actinobacillus actinomycetemcomitans and increased alamar fluorescence were noted . Metabolically active bacteria reduced alamar with the reduced dye found in the cell-free filtrate . Insignificant alamar reductions were noted in the absence of bacteria or by spent culture supernatants . The efficacy of mouthrinses with clinically proven antiplaque agents such as chlorhexidine or cetylpyridinium chloride were determined by alamar blue . In a model system with A . viscosus, triclosan dentifrices demonstrate a dose-dependent effect on bacteria . Human salivary bacteria demonstrate increasing alamar fluorescence with increasing plate counts . A clinical study examined the effects of rinsing with chlorhexidine or cetylpyridinium chloride mouthrinses in comparison with a placebo mouthrinse and water on salivary bacteria . Rinsing with chlorhexidine resulted in the least number of bacteria by alamar and plate count methods . In summary, the current study demonstrates the utility of alamar blue to examine the antimicrobial effects of oral care formulations in laboratory and clinical studies. Scand J Infect Dis, 2003, 35(9), 683 - 9 Intravenous immunoglobulin adjunctive therapy in sepsis, with special emphasis on severe invasive group A streptococcal infections; Norrby-Teglund A et al.; Immunotherapy targeted against microbial toxins and host mediators has been studied in many preclinical investigations and clinical trials of sepsis during the past 20 y . Intravenous immunoglobulin, including both monoclonal and polyclonal antibodies, represents one such immunotherapeutic strategy . Mononclonal antibodies directed against endotoxin or tumour necrosis factor-alpha have been tested extensively in clinical trials, but have so far failed to reveal a significant effect on mortality rates . Several studies have assessed the efficacy of polyclonal intravenous immunoglobulin (IVIG) in sepsis, with varying results . Although there are no conclusive data available to date to support the use of IVIG therapy in all sepsis cases, there are strong indications that certain defined septic subgroups, such as streptococcal toxic shock syndrome caused by group A streptococcus, will benefit from its use . This review briefly summarizes the clinical trials on IVIG therapy in sepsis, and describes in more detail the mechanistic actions of IVIG and the clinical data that support the use of IVIG as adjunctive therapy in severe invasive group A streptococcal infections. Scand J Infect Dis, 2003, 35(9), 647 - 52 Organ-specific models of Streptococcus pneumoniae disease; Orihuela CJ et al.; The variability of the course of infection by Streptococcus pneumoniae is well known but poorly understood . Most animal models of pneumonia, sepsis or meningitis have been forced to use site-specific bacterial inoculation to mimic localized human infection . This study examined the differences in the progression of disease-causing strains D39 (serotype 2), A66.1 (serotype 3) and TIGR4 (serotype 4) using isolates transformed with the Gram-positive lux transposon cassette, Tn4001 luxABCDE Km(r) . Expression of the lux operon results in bioluminescence, permitting the detection of the bacteria within a living animal while using a CCD camera . Mice infected intranasally with A66.1 developed only pneumonia, those challenged with D39 experienced high-grade sepsis, while TIGR4 infection resulted in low-grade pneumonia and bacteremia ultimately progressing to meningitis . Quantitative analysis of bacterial titers confirmed these patterns, which were consistent across different lineages of mice . Mice anesthetized with ketamine and xylazine developed more severe forms of the disease compared with isoflurane . These studies unambiguously characterize 3 distinct models of the natural course of pneumococcal infection . Mapping these models provides a framework for detailed molecular modeling of pneumococcal virulence determinants at specific stages of disease. Scand J Infect Dis, 2003, 35(9), 632 - 41 Proinflammatory activity of cell-wall constituents from gram-positive bacteria; Moreillon P et al.; Innate immunity reacts to conserved bacterial molecules . The outermost lipopolysaccharide (LPS) of Gram-negative organisms is highly inflammatory . It activates responsive cells via specific CD14 and toll-like receptor-4 (TLR4) surface receptor and co-receptors . Gram-positive bacteria do not contain LPS, but carry surface teichoic acids, lipoteichoic acids and peptidoglycan instead . Among these, the thick peptidoglycan is the most conserved . It also triggers cytokine release via CD14, but uses the TLR2 co-receptor instead of TLR4 used by LPS . Moreover, whole peptidoglycan is 1000-fold less active than LPS in a weight-to-weight ratio . This suggests either that it is not important for inflammation, or that only part of it is reactive while the rest acts as ballast . Biochemical dissection of Staphylococcus aureus and Streptococcus pneumoniae cell walls indicates that the second assumption is correct . Long, soluble peptidoglycan chains (approximately 125 kDa) are poorly active . Hydrolysing these chains to their minimal unit (2 sugars and a stem peptide) completely abrogates inflammation . Enzymatic dissection of the pneumococcal wall generated a mixture of highly active fragments, constituted of trimeric stem peptides, and poorly active fragments, constituted of simple monomers and dimers or highly polymerized structures . Hence, the optimal constraint for activation might be 3 cross-linked stem peptides . The importance of structural constraint was demonstrated in additional studies . For example, replacing the first L-alanine in the stem peptide with a D-alanine totally abrogated inflammation in experimental meningitis . Likewise, modifying the D-alanine decorations of lipoteichoic acids with L-alanine, or deacylating them from their diacylglycerol lipid anchor also decreased the inflammatory response . Thus, although considered as a broad-spectrum pattern-recognizing system, innate immunity can detect very subtle differences in Gram-positive walls . This high specificity underlines the importance of using well-characterized microbial material in investigating the system. J Bacteriol, 2003 Dec, 185(23), 6815 - 25 Factors contributing to hydrogen peroxide resistance in Streptococcus pneumoniae include pyruvate oxidase (SpxB) and avoidance of the toxic effects of the fenton reaction; Pericone CD et al.; Aerobic growth of Streptococcus pneumoniae results in production of amounts of hydrogen peroxide (H(2)O(2)) that may exceed 1 mM in the surrounding media . H(2)O(2) production by S . pneumoniae has been shown to kill or inhibit the growth of other respiratory tract flora, as well as to have cytotoxic effects on host cells and tissue . The mechanisms allowing S . pneumoniae, a catalase-deficient species, to survive endogenously generated concentrations of H(2)O(2) that are sufficient to kill other bacterial species is unknown . In the present study, pyruvate oxidase (SpxB), the enzyme responsible for endogenous H(2)O(2) production, was required for survival during exposure to high levels (20 mM) of exogenously added H(2)O(2) . Pretreatment with H(2)O(2) did not increase H(2)O(2) resistance in the mutant, suggesting that SpxB activity itself is required, rather than an H(2)O(2)-inducible pathway . SpxB mutants synthesized 85% less acetyl-phosphate, a potential source of ATP . During H(2)O(2) exposure, ATP levels decreased more rapidly in spxB mutants than in wild-type cells, suggesting that the increased killing of spxB mutants was due to more rapid ATP depletion . Together, these data support the hypothesis that S . pneumoniae SpxB contributes to an H(2)O(2)-resistant energy source that maintains viability during oxidative stress . Thus, SpxB is required for resistance to the toxic by-product of its own activity . Although H(2)O(2)-dependent hydroxyl radical production and the intracellular concentration of free iron were similar to that of Escherichia coli, killing by H(2)O(2) was unaffected by iron chelators, suggesting that S . pneumoniae has a novel mechanism to avoid the toxic effects of the Fenton reaction. Clin Microbiol Infect, 2003 Oct, 9(10), 1038 - 44 Prospective evaluation of blood cultures in a Turkish university hospital: epidemiology, microbiology and patient outcome; Esel D et al.; The aims of this prospective study were to: (1) determine the rate of blood culture contamination; (2) describe and compare the epidemiologic, clinical and microbiological characteristics of hospital- and community-acquired bloodstream infections; and (3) determine the mortality resulting from bloodstream infections . The rate of true bacteremia was 12.1%, and 10.7% of cultures were contaminated . Of the 567 episodes of bloodstream infection, 73.4% were hospital-acquired, and 26.6% were community-acquired . The most commonly isolated microorganisms were staphylococci (44%, methicillin resistant 69.4%), enterococci (15%) and Escherichia coli (12.5%) in hospital-acquired episodes, and Brucella spp . (21.9%), E . coli (19.2%) and Staphylococcus aureus (14.6%, methicillin resistant 9.1%) in community-acquired episodes . While the overall mortality rate was 25.4%, death attributable to bloodstream infections was 16.6% in hospital-acquired episodes and 13.9% in community-acquired episodes . The highest mortality occurred in patients with bacteremia due to Pseudomonas aeruginosa (37.5%) in hospital-acquired episodes, and in patients with bacteremia due to Streptococcus pneumoniae (50%) in community-acquired episodes . Underlying diseases, severity of illness, presence of bladder catheter, previous use of antibiotics, tracheal intubation and adequacy of treatment were found to be significantly associated with death. Clin Microbiol Infect, 2003 Oct, 9(10), 1031 - 4 Invasive properties of south Indian strains of Streptococcus pyogenes in a HEp-2 cell model; Gladstone P et al.; The objective of this study was to consider the invasive properties of Streptococcus pyogenes in human pharyngeal epithelial cells, and to correlate these with their clinical significance . Clinical isolates of S . pyogenes obtained from blood cultures over a period of 10 years, and throat and skin isolates from a community-based study, were used in this investigation . The S . pyogenes isolates were inoculated in HEp-2 cells and subsequently treated with antibiotics to kill the extracellular bacteria . The cells were then lyzed, and a colony count was carried out to check for invasion . The throat and skin isolates had 45.7%, 25.7% and 28.5% of low, intermediate and high invasion efficiencies, respectively, while 80%, 8.6% and 11.4% of the blood isolates had low, intermediate and high invasion efficiencies . We concluded that the throat and the skin isolates from superficial infections were more invasive than the blood isolates, which is an interesting and paradoxical feature. Clin Microbiol Infect, 2003 Aug, 9(8), 869 - 72 Streptococcus equi subspecies equi (Lancefield group C) meningitis in a child; Elsayed S et al.; We present a case of Streptococcus equi subsp . equi meningitis in a young boy . This case represents the first report in the literature of meningitis caused by this organism, as far as we know. Clin Microbiol Infect, 2003 Aug, 9(8), 832 - 8 Group B streptococcus colonization of Greek pregnant women and neonates: prevalence, risk factors and serotypes; Tsolia M et al.; OBJECTIVE: To evaluate the prevalence and risk factors of group B streptococcus (GBS) colonization among pregnant women and their neonates in Greece, and to examine the serotype distribution of the GBS strains isolated and their susceptibility to antibiotics . METHODS: A vaginal and a rectal swab were obtained from 1014 pregnant or parturient women followed at public and private hospitals in Athens and in a city of northern Greece . Cultures were also taken 24 h after birth from 428 neonates born to these women . RESULTS: The overall maternal and neonatal colonization rates were 6.6% and 2.4%, respectively . The vertical transmission rate was 22.5% . By logistic regression analysis, multiparity (>/=III) was associated with a lower colonization rate (odds ratio 4.4, 95% confidence interval 1.08-18.63) . In contrast with other studies, middle-class women followed privately were more frequently colonized (10%) than those followed at the public hospital (3.9%) (odds ratio 3.1, 95% confidence interval 1.83-5.42) . A higher number of prenatal visits was also associated with a higher colonization rate (change in true odds ratio when visits increased by one, 1.3; 95% confidence interval 1.14-1.60) . No association was found between colonization and maternal age, previous obstetric history, marital status, nationality, prematurity, Caesarean section, or infant birth weight . The most common serotypes were II (26.9%), III (22.4%), Ia (19%), Ib (12%), and V (9%) . A considerable proportion of the isolated strains was resistant to erythromycin (4.5%), clindamycin (6%), or both (6%) . CONCLUSION: The rate and risk factors of maternal and neonatal GBS colonization may vary in different communities . These rates, as well as the incidence of neonatal disease, need to be thoroughly evaluated in each country to allow the most appropriate preventive strategy to be selected. Clin Microbiol Infect, 2003 Sep, 9(9), 966 - 9 Development of macrolide resistance by ribosomal protein L4 mutation in Streptococcus pyogenes during miocamycin treatment of an eight-year-old Greek child with tonsillopharyngitis; Bozdogan B et al.; Streptococcus pyogenes isolates with the same pulsed-field patterns were recovered from the throat cultures of a child with tonsillopharyngitis before and after treatment with miocamycin, a 16-membered macrolide . The initial isolate was macrolide-susceptible, but the isolates after the treatment were resistant to 14 and 15-membered macrolides and had two amino acid (65WR66) deletions in ribosomal protein L4. Clin Microbiol Infect, 2003 Sep, 9(9), 944 - 8 Rhabdomyolysis as a result of Streptococcus pneumoniae: report of a case and review; Blanco JR et al.; We report the case of a patient who presented with RM associated with bacteremic pneumococcal pneumonia and review the literature on this condition. Clin Microbiol Infect, 2003 Sep, 9(9), 924 - 9 A common clone of erythromycin-resistant Streptococcus pneumoniae in Greece and the UK; Fotopoulou N et al.; OBJECTIVE: To investigate the possible genetic relationship among erythromycin-resistant Streptococcus pneumoniae strains isolated in Greece and the UK . METHODS: During 1995-97, 140 S . pneumoniae strains were isolated from clinical specimens submitted to the microbiology departments of the two main children's hospital in Athens . All erythromycin-resistant strains were further studied with respect to the presence of genes encoding for the two major mechanisms of macrolide resistance, their serotypes, and pulsed-field gel electrophoresis (PFGE) types, in comparison to a previously characterized UK erythromycin-resistant clone . RESULTS: Eleven of the 140 isolates (7.9%) were resistant to erythromycin; nine of these were susceptible to penicillin . Serotyping allocated seven, three and one isolates to serotypes 14, 19F and serogroup 6, respectively . The mefA gene was detected in seven isolates (five serotype 14 and two serotype 19F), ermB in two (one serotype 19F and the serogroup 6 isolate), whilst in the remaining two isolates no resistance gene could be detected by polymerase chain reaction (PCR) . Pulsed-field gel electrophoresis of genomic DNA showed that five Greek serotype 14 isolates belonged to the same chromosomal type as the serotype 14 erythromycin-resistant UK clone . CONCLUSIONS: The present study showed that erythromycin resistance among the S . pneumoniae isolates was mostly owing to the efflux mechanism and suggested a possible clonal spread of serotype 14 erythromycin-resistant S . pneumoniae strains between Greece and the UK. APMIS, 2003 Oct, 111(10), 945 - 50 Serologically indicated pneumococcal pneumonia in children: a population-based study in primary care settings; Heiskanen-Kosma T et al.; The aim of the study was to assess age-specific incidences of community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae and diagnosed serologically in a child population . The study was population-based, and prospective, and performed in primary health care settings . During a surveillance period of 12 months from 1981-1982, all pneumonia cases in a defined child population (57% urban residents) were registered prospectively . In total, 201 CAP cases were diagnosed (mean age 5.6 years; 57% boys; 58% urban residents) . S . pneumoniae etiology was studied by antibody and immune complex (IC) assays to C-polysacchride (C-PS), type-specific capsular polysaccharides (CPS), and to pneumolysin (Ply), in acute and convalescent sera . Serologic evidence of S . pneumoniae etiology was indicated in 57(28%) cases, 35(61%) being mixed infections with other microbes . The distribution of pneumococcal cases was 44%, 30% and 26% in the three 5-year age groups, respectively . There were 33 (58%) males and 34 (60%) urban residents . In total, 26 cases were identified by antibody assays and 35 cases by IC assays, 26/35 being positive in acute sera . Responses to C-PS, CPS and Ply, when antibody and IC results are combined, were found equally often in 23-25 cases . The total annual incidence of pediatric S . pneumoniae CAP was 6.4/1000/year . S . pneumoniae etiology was found in 28% of the children and was similar at all ages . The incidence of pneumococcal CAP was assessed for the first time, being high (19/1000/year) in 0- to 4-year-old urban boys and rather stable (5-9/1000/year) in all other groups by age, sex and residence. Aliment Pharmacol Ther, 2003 Nov 15, 18(10), 949 - 62 Review article: oral ulceration--aetiopathogenesis, clinical diagnosis and management in the gastrointestinal clinic; Field EA et al.; Oral ulceration is a common complaint of patients attending out-patient clinics . The aim of this review is to provide the gastroenterologist with a differential diagnosis of oral ulceration, and a practical guide for the management of recurrent aphthous stomatitis, including topical and systemic therapy . The association of recurrent aphthous stomatitis with Behcet's disease and other systemic disorders, including coeliac disease, is discussed . Recent evidence concerning the immunopathogenesis of Behcet's disease is reviewed, including renewed interest in the role of Streptococcus sanguis and possible infectious triggering of an inappropriate immunoinflammatory response, resulting in tissue damage . The efficacy and limitations of conventional treatment for this mutisystem disorder are outlined together with the potential role of novel biological agents, such as anti-tumour necrosis factor-alpha therapy . Oral ulceration, as a manifestation of inflammatory bowel disease and a complication of drug therapy, is described . Guidance is given concerning indications for referral of patients with oral ulceration to an oral physician/surgeon for further investigations, including biopsy if appropriate. Clin Infect Dis, 2003 Dec 1, 37(11), 1534 - 40 Epub 2003 Nov 06. Decreased serum opsonic activity against Streptococcus pneumoniae in human immunodeficiency virus-infected Ugandan adults; Takahashi H et al.; Type-specific immunoglobulin G (IgG) to pneumococcal capsular polysaccharide (CPS) and opsonic activity against Streptococcus pneumoniae were evaluated in serum samples from 36 Ugandan adults with community-acquired pneumonia and 58 asymptomatic Ugandan adults with or without human immunodeficiency virus type 1 (HIV-1) infection . The levels of serum IgG to CPS were significantly higher in HIV-1-infected subjects than in HIV-uninfected subjects . Serum samples from HIV-1-infected subjects that had lower IgG titers demonstrated higher opsonic activity against type 3 (titers of 7) and type 9 (titers of 7-11) pneumococcal strains . Plasma HIV-1 load also correlated inversely with serum opsonic activity against these strains, and peripheral blood CD4+ lymphocyte numbers also tended to correlate with serum opsonic activity in asymptomatic HIV-1-infected adults . Our findings suggest that the opsonic activity of type-specific IgG is impaired in the serum of HIV-1-infected African adults, which may expose them to a serious risk of invasive pneumococcal infections. Clin Infect Dis, 2003 Dec 1, 37(11), 1481 - 9 Epub 2003 Nov 07. Clinical presentation and bacteriologic analysis of infected human bites in patients presenting to emergency departments; Talan DA et al.; Previous studies of infected human bites have been limited by small numbers of patients and suboptimal microbiologic methodology . We conducted a multicenter prospective study of 50 patients with infected human bites . Seventy percent of the patients and assailants were young adult men . Fifty-six percent of injuries were clenched-fist injuries and 44% were occlusional bites . Most injuries were to the hands . Fifty-four percent of patients were hospitalized . The median number of isolates per wound culture was 4 (3 aerobes and 1 anaerobe); aerobes and anaerobes were isolated from 54% of wounds, aerobes alone were isolated from 44%, and anaerobes alone were isolated from 2% . Isolates included Streptococcus anginosus (52%), Staphylococcus aureus (30%), Eikenella corrodens (30%), Fusobacterium nucleatum (32%), and Prevotella melaninogenica (22%) . Candida species were found in 8% . Fusobacterium, Peptostreptococcus, and Candida species were isolated more frequently from occlusional bites than from clenched-fist injuries . Many strains of Prevotella and S . aureus were beta-lactamase producers . Amoxicillin-clavulanic acid and moxifloxacin demonstrated excellent in vitro activity against common isolates. J Med Microbiol, 2003 Dec, 52(Pt 12), 1047 - 58 Using cpsA-cpsB sequence polymorphisms and serotype-/group-specific PCR to predict 51 Streptococcus pneumoniae capsular serotypes; Kong F et al.; Streptococcus pneumoniae polysaccharide and protein-conjugate vaccines are available against the most commonly isolated pneumococcal serotypes . Ongoing surveillance of invasive pneumococcal disease is needed in order to monitor changes in distribution of serotypes . Based on previously published sequences of capsular polysaccharide synthesis (cps) gene clusters of 16 pneumococcal serotypes, a molecular capsular typing (MCT) system has been developed, based on a combination of partial cpsA-cpsB sequencing and serotype- or serogroup-specific PCR, targeting the genes wzy and wzx (except for serotype 3) . In this study, 151 S . pneumoniae isolates of known serotype (representing 51 serotypes) and 276 recent clinical isolates were used to develop MCT and compare it with conventional serotyping (CS) (total 427 isolates) . On the basis of 376 heterogeneity sites in the cpsA-cpsB region, 89 sequence types (ST) were identified, of which 76 corresponded to a single serotype and 11 contained two serotypes . The correct serotypes in two of the latter (10A-23F-g and 23F-23A) were identified using serotype 23F-specific PCR . Limited CS was required for 92 (22 %) isolates to distinguish between the two serotypes in the nine other mixed ST (6A-6B-g, 6A-6B-q, 15B-22F, 33F-33A, 17F-35B, 18B-18C, 13-20, 25F-38, 31-42) . MCT is a specific, objective and practical method that can predict the serotype of most S . pneumoniae isolates; it will facilitate epidemiological studies . Further study of the relationship between MCT and CS is needed in order to improve our understanding of serotype differentiation and to improve MCT methods further. J Antimicrob Chemother, 2003 Dec, 52(6), 967 - 73 Epub 2003 Nov 12. Phage lytic enzymes as therapy for antibiotic-resistant Streptococcus pneumoniae infection in a murine sepsis model; Jado I et al.; OBJECTIVES: Phage-coded lysins, i.e . murein hydrolases, are enzymes that destroy the cell wall of bacteria . A rapid killing of Streptococcus pneumoniae in the nasopharynx of mice has been described recently using a phage-coded murein hydrolase (enzybiotic) . The in vivo effects of a dose-ranging treatment, using either of the phage-coded lytic enzymes Cpl-1 lysozyme or the Pal amidase, have been investigated here in a murine sepsis model . METHODS: Purified Pal amidase and/or Cpl-1 lysozyme were used alone or in combination . These enzymes were injected intraperitoneally at different times after challenge with 5 x 10(7) cfu of a type 6B, antibiotic-resistant S . pneumoniae clinical isolate . RESULTS: Animals challenged with 5 x 10(7) cfu of this strain alone died within 72 h, whereas a single intraperitoneal injection of Cpl-1 or Pal (200 microg; 1100 U) administered 1 h after the bacterial challenge was sufficient to effectively protect the mice, according to unpaired t-test (P<0.0001) . Bacteraemia in unprotected mice reached colony counts >10(7) cfu/mL, whereas the mean colony count in lysin-protected animals was <10(6) cfu/mL over time and ultimately became undetectable . Interestingly, a synergic effect in vivo was observed with the combined use of 2.5 microg each of Cpl-1 and Pal . CONCLUSIONS: Our findings suggest strongly that phage lysins protect animals from bacteraemia and death . Moreover, the simultaneous attack of the pneumococcal peptidoglycan by a lysozyme and an amidase leads to a remarkable effect through enhanced destruction of the bacterial cell wall . The benefits of therapy with enzybiotics against pneumococcus reported here might warrant the examination of alternative strategies for the treatment of diseases caused by clinically relevant pathogens. Ann Clin Microbiol Antimicrob . 2003 Nov 7;2(1):9. Recurrent pneumococcal meningitis in a splenectomised HIV-infected patient; Morand PC et al.; BACKGROUND: Streptococcus pneumoniae is a major cause of human disease, especially in pre-school children and elderly people, as well as in special risk groups such as asplenic, antibody deficient patients, or presenting disruption of natural barriers . The occurrence of pneumococcal disease has increased with the onset of the HIV epidemic and the emergence of drug-resistance . CASE PRESENTATION: We report the case of an HIV-1-infected patient who experienced three episodes of recurrent pneumococcal meningitis over a 4-year period, despite chemoprophylaxis and capsular vaccination . CONCLUSIONS: Efficacy of anti-pneumococcal chemoprophylaxis and vaccination in HIV-infected patients are discussed in the light of this particular case. Curr Opin Allergy Clin Immunol, 2003 Dec, 3(6), 451 - 9 Primary immunodeficiencies associated with pneumococcal disease; Picard C et al.; PURPOSE OF REVIEW: Streptococcus pneumoniae may cause disease in patients with a variety of primary immunodeficiencies . However, no previous review has dealt with the issue of which primary immunodeficiencies predispose affected individuals to pneumococcal disease . We thus reviewed the medical literature on cases of S . pneumoniae infection in patients with primary immunodeficiency diseases, with a particular emphasis on invasive pneumococcal disease . RECENT FINDINGS: Primary immunodeficiency diseases comprise over 100 conditions, each associated with a variety of infections . Patients at high risk for pneumococcal disease include most if not all B-cell defects (whether due to an intrinsic B-cell anomaly or an impaired T-cell help), deficiencies of early components of the classical pathway of complement and C3 deficiency, congenital asplenia, anhidrotic ectodermal dysplasia with immunodeficiency (caused by impaired NF-kappaB activation), and interleukin-1 receptor associated kinase-4 deficiency . Patients with other complement deficiencies (alternative and third pathway) and hyperimmunoglobulin E syndrome show a lower risk, whereas patients with other known primary immunodeficiencies, such as phagocytic disorders, do not appear to be particularly vulnerable to S . pneumoniae . SUMMARY: Antibody- and complement-mediated opsonization, splenic macrophages and interleukin-1 receptor associated kinase-4- and nuclear factor kappaB-mediated immune responses are crucial for protective immunity to S . pneumoniae . This information is useful, not only in increasing our understanding of human immunity to S . pneumoniae, but also in the diagnostic investigation of patients with pneumococcal disease. J Biol Chem, 2004 Jan 30, 279(5), 3375 - 81 Epub 2003 Nov 11. Improving protein pharmacokinetics by genetic fusion to simple amino acid sequences; Alvarez P et al.; The role of primary amino acid sequences in protein pharmacokinetics, an issue of relevance in both basic knowledge and biotechnology, was addressed here using as a starting point two repetitive antigens from the hemoflagellate Trypanosoma cruzi that are known to stabilize their associated proteins in the bloodstream . A major drawback to their pharmacological application is that these repetitive sequences are highly immunogenic, being therefore the deletion of this characteristic desirable . Based on sequence homology and epitope mapping analyses, an artificial repetitive sequence (PSTAD) was engineered . This motif was tested by genetic fusion to the C terminus of both the trypanosomal trans-sialidase and the rat tyrosine aminotransferase and found to produce a 4.5-6-fold increase in the half-life of the associated proteins in blood while displaying significantly lower immunogenicity . Residues involved in the stabilizing properties of the novel peptide were mapped by a site-directed mutagenesis approach, allowing us to successfully identify another two motifs . Searching databases for sequences displaying some homology, embedded in proline frameworks and associated to shed virulence factors from unrelated microorganisms, resulted in the identification of four other protein extensions . Remarkably, three of them (from Streptococcus pneumoniae, Actinomyces viscosus, and Escherichia coli) revealed similar pharmacokinetic features, suggesting therefore an analogous evolutionarily acquired mechanism to ensure the biodistribution of their corresponding proteins . Our findings indicate that the insertion of defined motifs into a proline-rich framework constitutes a suitable alternative to construct a chimeric protein with extended half-life in blood. Arch Dis Child, 2003 Nov, 88(11), 1009 - 14 Empyema thoracis: a 10-year comparative review of hospitalised children from south Asia; Baranwal AK et al.; AIMS: To study the clinical and microbial profile of childhood empyema in South Asia and to identify the changes over the past three decades . METHODS: A total of 265 children (aged 1 month to 12 years) with empyema admitted to the Advanced Pediatric Center, PGIMER, Chandigarh, India in 1989-98, were reviewed retrospectively . RESULTS AND CONCLUSIONS: One third of children were under 5 . Culture positivity had decreased significantly (48% v 75%) over the years . Staphylococcus aureus continues to be the commonest (77%) aetiological agent; clustering was seen during hot and humid months (46%) . Culture positive Streptococcus pneumoniae cases also decreased (9% v 27%); all were seen during the winter and spring season . Gram negative rods grew in more patients (11% v 7%) . Community acquired methicillin resistant S aureus (MRSA) was isolated in three patients . Most children (93%) were treated with parenteral cloxacillin and an aminoglycoside . Tube drainage (TD) was used in 92% of fibropurulent cases, and was successful in 79% . Of 48 patients with failed TD, 12 needed decortication; limited thoracotomy was sufficient in the remaining 36 . Surgery was mainly required by children with persistent pleural sepsis after 10 days of TD . Delaying surgery until 14 days had a significantly higher potential of requiring decortication . Early change to oral antibiotics (after 1-2 weeks of parenteral therapy) reduced the hospital stay significantly (17+7 v 23+7 days) without compromising long term outcome . Twenty two patients presenting late in the chronic stage underwent decortication at admission. J Ethnopharmacol, 2003 Dec, 89(2-3), 301 - 3 Antibacterial activity of Leontopodium alpinum (Edelweiss); Dobner MJ et al.; Extracts and individual constituents of Leontopodium alpinum Cass . (Asteraceae) were tested for their antimicrobial activity in two different assays . Extracts were screened in agar diffusion assays, whereas the minimum inhibitory concentrations (MIC) of single compounds were determined by the microbroth dilution method according to NCCLS criteria . Significant antimicrobial activities were found against various strains of Enterococcus faecium, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae, and Streptococcus pyogenes strains . These results support the ethnomedicinal use of Leontopodium alpinum for the treatment of respiratory and abdominal disorders. Eur J Pediatr, 2004 Jan, 163(1), 19 - 21 Epub 2003 Nov 11. Group A streptococcal meningitis as a complication of an infected capillary haemangioma; Rezvani M et al.; We report a case of group A streptococcal meningitis in an infant resulting from an infected capillary haemangioma . The child suffered significant morbidity including cerebral infarction, epilepsy, and developmental delay . Treatment of infected capillary haemangiomas remains controversial and inconsistent . CONCLUSION: Our experience of this infant, resulting in profound neurological morbidity suggests that group A Streptococcus can be a virulent organism in the young child and that capillary haemangiomas must be treated aggressively at the first sign of infection. Proc Natl Acad Sci U S A, 2003 Nov 25, 100(24), 14363 - 7 Epub 2003 Nov 10. Pneumococcal carriage results in ganglioside-mediated olfactory tissue infection; van Ginkel FW et al.; Streptococcus pneumoniae cause considerable morbidity and mortality, with persistent neurological sequelae, particularly in young children and the elderly . It is widely assumed that carriage occurs through direct mucosal colonization from the environment whereas meningitis results from invasion from the blood . However, the results of published studies can be interpreted that pneumococci may enter the brain directly from the nasal cavity by axonal transport through olfactory nerves . This hypothesis is based on findings that (i) teichoic acid of the pneumococcal cell wall interact with gangliosides (GLS), (ii) the interaction of GLS with cholera toxin leads to axonal transport through the olfactory nerves into the brain, and (iii) viruses enter the brain through axonal transport into olfactory nerves . After nasal inoculation, we observe high numbers of pneumococci in nasal washes and the olfactory nerves and epithelium . Significant numbers of pneumococci also infected the olfactory bulbs, brain, and the trigeminal ganglia . The absence of bacteremia in this model makes it unlikely that the bacteria entered the brain from the blood stream . Recovery of colony-forming units from the brain, lungs, olfactory nerves, and epithelium and nasal washes was inhibited by incubating pneumococci with GLS before nasal inoculation . These findings, confirmed by PCR and immunohistochemistry, support a GLS-mediated process of infection and are consistent with pneumococci reaching the brain through retrograde axonal transport. Minerva Pediatr, 2003 Oct, 55(5), 439 - 45 {Nasopharyngeal colonization of Streptococcus pneumoniae in healthy children: percentage of carriers, serotypes distribution and antibiotic resistance}; Schettini F Jr et al.; AIM: The nasopharyngeal carriage of Streptococcus pneumoniae is an important risk factor for pneumococcal diseases . Data regarding prevalence and serotype distribution of this pathogen are lacking in our population . METHODS: Experimental design: longitudinal observational cohort study . Setting: healthy children aged 1-7 years attending day-care centers and schools of a district of a Southern Italy city . Measures: the nasopharyngeal colonization rate of Streptococcus pneumoniae as well as its antibiotic susceptibility was determined . RESULTS: Of 317 nasopharyngeal cultures obtained, 18.29% of the cultures were positive for Streptococcus pneumoniae; 60.34% of the isolates were serotypes 19A, 19F, 14, 6B, or 23F; 8.62% of the strains were intermediately resistant to penicillin . Erythromycin-resistance was observed in 65.51% of the micro-organisms isolated and particularly serotypes 19, 14, and 6 were more erythromycin-resistant than organisms of other serotypes . Co-trimoxazole resistance was detected in 17.24% of the strains . All the strains resulted uniformly susceptible to cefotaxime and ceftriaxone . CONCLUSION: The high rate of nasopharyngeal carriage of Streptococcus pneumoniae, along with the resistance to antibiotics widely used in the community, suggests the importance of an epidemiological surveillance as well as the application of new vaccine strategies. Clin Diagn Lab Immunol, 2003 Nov, 10(6), 1019 - 24 Multilaboratory evaluation of a viability assay for measurement of opsonophagocytic antibodies specific to the capsular polysaccharides of Streptococcus pneumoniae; Romero-Steiner S et al.; Opsonophagocytosis is a correlate of protection that measures the functional activity of vaccine-induced antibodies . A standardized opsonophagocytosis assay (OPA) should be used as part of the evaluation of current and future pneumococcal (Pnc) polysaccharide (Ps)-based vaccines . We enrolled five laboratories to evaluate a previously standardized viability OPA . Each laboratory was provided with a detailed OPA protocol, seven target Pnc strains (serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F), two quality control sera and 12 paired sera (blinded) from adult donors who received one dose of the 23-valent Pnc Ps vaccine . Laboratories sent their results to the Centers for Disease Control and Prevention for analysis . Sera were tested in duplicate (single run), and the results were averaged to yield a single OPA titer (> or = 50% killing) for each serum sample . The percentage of sera within one or two dilutions of the calculated median OPA titer was determined for each laboratory and for each serotype . In general, laboratories were capable of detecting OPA titers within one or two dilutions of the median for at least 75 and 88%, respectively, of the sera tested . The level of agreement with the median OPA titers varied depending on the participating laboratory (overall agreement = 0.8 {99% confidence interval = 0.75 to 0.85}) . All OPA median titers reported for quality control sera were within one dilution of the expected titer . We conclude that this OPA can be done in multiple laboratories with a high degree of interlaboratory reproducibility. Scand J Infect Dis, 2003, 35(10), 757 - 9 Different polymerase chain reaction-based analyses for culture-negative endocarditis caused by Streptococcus pneumoniae; Voldstedlund M et al.; Two cases of culture-negative infective endocarditis caused by Streptococcus pneumoniae are presented . Conventional bacteriological methods were compared with 2 different polymerase chain reaction (PCR)-based analyses: PCR amplification with primers specific for S . pneumoniae, and broad-range PCR followed by sequencing . PCR-based analyses can be a valuable supplement to traditional bacteriological analyses. Immunogenetics, 2003 Dec, 55(9), 629 - 32 Epub 2003 Nov 06. Alleles carried at positions -819 and -592 of the IL10 promoter affect transcription following stimulation of peripheral blood cells with Streptococcus pneumoniae; Temple SE et al.; IL-10 inhibits the production of many pro-inflammatory cytokines . Polymorphisms in the IL10 gene promoter at positions -1082G-->A, -819C-->T and -592C-->A occur as three haplotypes, ATA, GCC and ACC . These influence several infectious and inflammatory diseases including community-acquired pneumonia, where a role for IL-10 is suggested by fluctuations in plasma levels of the cytokine . However, the effects of the haplotypes on IL-10 production are unclear . We stimulated peripheral blood mononuclear cells (PBMC) from at least five individuals homozygous for each of the three haplotypes with lipopolysaccharide (LPS, 10 microg/ml) or heat-killed Streptococcus pneumoniae (10(7)cfu/ml) and measured IL-10 mRNA by RT-PCR . Following S . pneumoniae stimulation, PBMC with the ATA haplotype had higher IL-10 mRNA levels than those with the GCC haplotype at 4 h (independent t-test; P=0.024), or the ACC haplotype at 4 h ( P<0.0001) and 8 h ( P=0.007) . Following LPS stimulation, IL-10 mRNA levels were not significantly influenced by the IL10 haplotype, but similar trends were observed, consistent with the variable outcome of published studies . The results suggest that the -819 and/or -592 alleles affect transcription. J Clin Microbiol, 2003 Nov, 41(11), 5288 - 90 Infections in international pregnancy study: performance of the optical immunoassay test for detection of group B streptococcus; Thinkhamrop J et al.; We evaluated the Strep B optical immunoassay (OIA; ThermoBiostar, Inc.) for detecting light and heavy group B streptococcus colonization in 1,306 pregnant women . The women were examined at 20 to 32 weeks gestation and were from six countries . Compared to culture, the sensitivity and specificity of OIA were 13.3 and 98.4%, respectively, for light colonization and 41.5 and 97.7%, respectively, for heavy colonization. J Clin Microbiol, 2003 Nov, 41(11), 5285 - 7 Prevalence of group A streptococcal carriers in asymptomatic children and clonal relatedness among isolates in Malatya, Turkey; Durmaz R et al.; In our study, the prevalence of nasopharyngeal Streptococcus pyogenes was 130 (14.3%) of 909 healthy children . Isolates were found to be susceptible to all antibiotics tested . Pulsed-field gel electrophoresis and arbitrarily primed PCR revealed that 34 (32.4%) of the 105 isolates and 41 (40.6%) of the 101 isolates typed, respectively, were clonally indistinguishable. J Clin Microbiol, 2003 Nov, 41(11), 5282 - 4 Bacitracin-resistant clone of Streptococcus pyogenes isolated from pharyngitis patients in Belgium; Malhotra-Kumar S et al.; We report 16 bacitracin-resistant Streptococcus pyogenes isolates recovered from pharyngitis patients in Belgium, 14 of which belonged to a particular emm type (emm28) . All 16 isolates were constitutively resistant to macrolides and carried erm(B) . The emergence of a bacitracin-resistant S . pyogenes clone raises questions about the continued reliability of bacitracin susceptibility testing for S . pyogenes identification. J Clin Microbiol, 2003 Nov, 41(11), 4966 - 70 Geographic distribution and clonal diversity of Streptococcus pneumoniae serotype 1 isolates; Brueggemann AB et al.; Serotype 1 pneumococci are a major cause of serious disease and have been associated with outbreaks but are rarely carried . The high attack rate and lack of coverage of this serotype by the heptavalent conjugate vaccine prompted the characterization of a geographically diverse collection of 166 serotype 1 isolates from recent cases of invasive disease . The isolates were resolved by multilocus sequence typing into 16 clones, which clustered into three major lineages with very different geographic distributions . Lineage A isolates were exclusively from Europe and North America, lineage B isolates were predominantly from Africa and Israel, and lineage C isolates were mainly from Chile . There was no clear association between the presence of individual clones within a country and the prevalence of serotype 1 disease. Laryngoscope, 2003 Nov, 113(11), 2047 - 51 Acute otitis media develops in the rat after intranasal challenge of Streptococcus pneumoniae; Eriksson PO et al.; OBJECTIVES/HYPOTHESIS: The rat is a frequently used animal model for middle ear research . To date, acute otitis media (AOM) has been evoked after instillation of bacteria directly into the middle ear cavity or after traumatizing the tympanic membrane . The purpose of the study was to examine whether, with an intact tympanic membrane and middle ear cavity, intranasally deposited bacteria cause AOM and how tympanic membrane stimulation influences this procedure . STUDY DESIGN: In vivo, murine model . METHODS: In a rat model, Streptococcus pneumoniae, type 3, was intranasally inoculated for 5 consecutive days . The tympanic membrane was treated with saline or with compound 48/80 or was left untreated . The development of AOM was evaluated by otomicroscopy, light microscopy, and middle ear culture . RESULTS: Ninety percent of the ears developed AOM . However, when the tympanic membranes were treated with saline or compound 48/80, only 40% and 57%, respectively, developed AOM . In all, 23 of 40 ears developed AOM and 20 ears showed growth of bacteria . CONCLUSION: Repeated intranasal deposition of S . pneumoniae, type 3, causes AOM in the rat . The development of AOM can be influenced by tympanic membrane stimulation. Int J Antimicrob Agents, 2003 Nov, 22(5), 541 - 4 Evolution of penicillin and erythromycin co-resistance in Streptococcus pneumoniae in Spain; Garcia-Rey C et al.; The temporal dynamics of penicillin and erythromycin co-resistance in Streptococcus pneumoniae based on two extensive multicentre Spanish surveillance SAUCE studies (1996-1997 and 1998-1999) is presented . Erythromycin resistance among penicillin non-susceptible isolates seems to have reached a limit as evidenced by a null increase between the two surveys, whereas it is growing among penicillin-susceptible pneumococci. Z Geburtshilfe Neonatol, 2003 Sep-Oct, 207(5), 186 - 9 {Severe late-onset group B streptococcal infection . A case report}; Haase R et al.; BACKGROUND: Group B Streptococcus (GBS) is a well-known cause of neonatal pneumonia, sepsis and meningitis . Peripartal antibiotic prophylaxis for early-onset GBS infection is in routine use since the beginning of the last decade, but strategies for effective prevention of late-onset GBS infections are still lacking . CASE REPORT: Few hours after discharge from a non-local maternity ward a 3-week-old boy was admitted to our hospital because of GBS meningitis with necrotizing encephalomalacia . Maternal mastitis, not a disease of the baby, had led to the first admission . DISCUSSION: Case history and negative maternal swabs and cultures for GBS led to the hypothesis of nosocomial infection . Screening and risk based peripartal antibiotic prophylaxis, better monitoring and improved therapeutic modalities have reduced the incidence and mortality of early-onset GBS infections, but peripartal prophylaxis failed to influence late-onset GBS infections . Up to 40 % of infants with late-onset meningitis develop neurological sequelae . Maternal vaccination with multivalent conjugate vaccines against GBS is a new strategy which may lead to passive protection of the infant . Further studies to examine the efficacy of vaccines are in progress. J Clin Pathol, 2003 Nov, 56(11), 879 - 81 Corticosteroids as adjunctive treatment in Austrian's syndrome (pneumococcal endocarditis, meningitis, and pneumonia): report of two cases and review of the literature; du Cheyron D et al.; This report describes two cases of Osler's triad of pneumonia, meningitis, and endocarditis, as a result of Streptococcus pneumoniae infection, also called Austrian's syndrome . In the first patient, a 51 year old non-alcoholic man, the aortic valve was affected and needed to be replaced in an emergency operation . The mitral valve was affected in a 70 year old woman without underlying disease, who only benefited from medical treatment . Both patients received corticosteroids, either dexamethasone followed by low doses of hydrocortisone and fludrocortisone, or only hydrocortisone and fludrocortisone, at the onset of the illness, and their outcome was favourable . These case reports focus on the presentation, prognosis, and therapeutic options for this severe syndrome. Nippon Ganka Gakkai Zasshi, 2003 Oct, 107(10), 590 - 6 {Vitrectomy for endophthalmitis after cataract surgery}; Oshitari K et al.; PURPOSE: To identify risk factors of poor visual outcome with vitrectomy for early-onset endophthalmitis after cataract surgery . PATIENTS AND METHODS: Clinical records of 29 consecutive eyes with endophthalmitis developing within 6 weeks after cataract surgery and that underwent therapeutic vitrectomy between June 1996 and April 2001 were retrospectively reviewed . Twenty-two of the eyes received intravitreal injections of vancomycin and ceftazidime at the time of vitrectomy, and all patients received intravenous antibiotics . Eyes were divided into two groups; group A consisted of 22 eyes with a final visual acuity of 0.2 or greater, and group B consisted of 7 eyes with a final visual acuity of less than 0.2 . RESULTS: Fifteen eyes (52%) in group A achieved a visual acuity of 0.5 or better and 8(28%) achieved a visual acuity of 1.0, while 4 eyes in group B developed phthisis bulbi . For eyes with a preoperative visual acuity of hand motions or worse, there was no correlation between final visual acuity and preoperative visual acuity . The overall culture-positive rate was 57% . In group A, methicillin-resistant Staphylococcus epidermidis was identified in 6 eyes, methicillin-resistant Staphylococcus aureus (MRSA) in 3 eyes and enterococcus in 2 eyes . In group B, alpha-hemolytic streptococcus (AHS) was identified in 4 eyes, aspergillus in 1 eye, and MRSA in 1 eye . All isolates were sensitive to vancomycin with the exception of the aspergillus . AHS infection appeared to be associated with wound failure from the initial cataract surgery and a poor visual outcome . Among 3 of the eyes that developed phthisis bulbi, intravitreal injection of antibiotics was not performed . CONCLUSION: Early vitrectomy and intravitreal injection of vancomycin may improve visual outcomes, but infection with AHS may be associated with cataract surgery wound failure and poor visual outcomes. Arch Oral Biol, 2003 Dec, 48(12), 815 - 24 A 100 kDa vanadate and lanzoprazole-sensitive ATPase from Streptococcus mutans membrane; Magalhaes PP et al.; The cariogenic potential of Streptococcus mutans is due to the production of organic acids derived from energy metabolism, which implies the need of mechanisms for the organism to tolerate this acidic environment . The F(1)F(o)-ATPase is generally considered as the main enzyme responsible for cytoplasmic proton extrusion, but mutations that resulted in a 50% reduction in F(1)F(o)-ATPase activity in S . mutans still allowed the micro-organism to grow and extrude acid, keeping the intracellular pH one pH unit above the extracellular ambient . This finding suggests the existence of other enzymatic (or cellular) mechanisms that keep the cytosolic pH neutral during micro-organism growth . This paper describes a membrane protein in S . mutans, with a molecular weight of 100 kDa, which exhibits ATPase activity inhibited by classic inhibitors of P-type ATPases (orthovanadate) and H(+),K(+)-ATPase (lanzoprazole), has an optimum pH comparable to other H(+)-ATPases and undergoes phosphorylation during the catalytic reaction, like that of H(+)-ATPases described in yeast and plant plasma membrane . Together, these results strongly suggest that the enzyme we describe here is a P-type H(+)-ATPase or H(+),ion-ATPase that can act in association with F(1)F(o)-ATPase during the growth of the S . mutans. Pediatrics, 2003 Nov, 112(5), 1095 - 102 Streptococcus pneumoniae infections in the neonate; Hoffman JA et al.; OBJECTIVE: Streptococcus pneumoniae infections in the neonate (SPIN) are relatively unusual events (1%-11% of neonatal sepsis) but are associated with substantial morbidity and mortality . Previous reports suggest that invasive SPIN is associated with prolonged rupture of membranes, maternal colonization/illness, prematurity, early-onset pneumonia presentation (<72 hours), and high mortality (50%) . The aim of this study was to review the current epidemiology and clinical course of SPIN . METHODS: The US Pediatric Multicenter Pneumococcal Surveillance Group has been prospectively monitoring S pneumoniae infections since 1993 in 8 children's hospitals . For this report, data were gathered retrospectively from the charts of neonates who were 30 days of age and younger and had SPIN from September 1993 to February 2001 . All pneumococcal isolates were sent to a central laboratory for serogrouping/typing and susceptibility testing . RESULTS: Twenty-nine cases of SPIN were identified from a total of 4428 episodes of S pneumoniae infection in children . Sixty-six percent were male, and 55% were white; the mean age was 18.1 day (+/-8.2) . Ninety percent of infants were >or=38 weeks' gestation . Two mothers had bacterial infections at delivery; 1 had S pneumoniae isolated from both blood and cervix, and 1 had clinical amnionitis . The primary diagnoses in the neonates were bacteremia (8), meningitis (8), bacteremic pneumonia (4), septic arthritis/osteomyelitis (1), and otitis media (8) . Thirty percent of infants with invasive SPIN presented with leukopenia/neutropenia, but this did not predict poor outcome . The infecting pneumococcal serogroups were 19 (32%); 9 (18%); 3 and 18 (11% each); 1, 6, and 14 (7% each); and 5 and 12 (3.5% each) . Twenty-six percent of invasive neonatal infections were caused by serogroups 1, 3, 5, and 12, which are not contained in the heptavalent pneumococcal vaccine . In contrast, 6% of invasive nonneonatal disease was caused by these same nonvaccine serogroups . Susceptibility testing demonstrated that 21.4% of isolates were penicillin nonsusceptible and 3.6% were ceftriaxone nonsusceptible . Three (14.3%) neonates with invasive SPIN died; all deaths occurred within 36 hours of presentation . Deaths did not appear to be related to pneumococcal serogroup or susceptibilities . CONCLUSIONS: Compared with previous studies of neonates with pneumococcal infection, this series showed that infants with SPIN were usually 2 to 3 weeks of age at presentation; likely to be full term; and ill with pneumonia, meningitis, and otitis media . This late-onset presentation was associated with an overall mortality rate of 10.3% (14.3% for invasive disease). J Bacteriol, 2003 Nov, 185(22), 6640 - 7 MgrA, an orthologue of Mga, Acts as a transcriptional repressor of the genes within the rlrA pathogenicity islet in Streptococcus pneumoniae; Hemsley C et al.; Streptococcus pneumoniae normally resides in the human nasopharynx in a nondisease state . In response to unknown triggers this organism can descend to the lower respiratory tract and/or invade the bloodstream . Regulation and activation of virulence genes play essential roles in this process of disease development . Characterization of S . pneumoniae regulatory networks has been a recent area of interest, but despite inroads little is known about regulation of virulence genes in this pathogen . A putative transcriptional regulator in S . pneumoniae, mgrA, which exhibits homology to the virulence gene activator mga of group A streptococcus, was previously identified as a regulator that is required for development of pneumonia in a murine model . In this study we confirmed that mgrA plays a role in both nasopharyngeal carriage and pneumonia . Transcriptional profiling by microarray technology was used to show that mgrA acts as a repressor of the previously characterized rlrA pathogenicity islet . This is manifested phenotypically by a decrease in adherence to epithelial cells in tissue culture since the rlrA pathogenicity islet contains genes mediating adherence. J Bacteriol, 2003 Nov, 185(22), 6592 - 9 MtaR, a regulator of methionine transport, is critical for survival of group B streptococcus in vivo; Shelver D et al.; The group B streptococcus (GBS) is an important human pathogen that infects newborns as well as adults . GBS also provides a model system for studying adaptation to different host environments due to its ability to survive in a variety of sites within the host . In this study, we have characterized a transcription factor, MtaR, that is essential for the ability of GBS to survive in vivo . An isogenic strain bearing a kanamycin insertion in mtaR was attenuated for survival in a neonatal-rat model of sepsis . The mtaR mutant grew poorly in human plasma, suggesting that its utilization of plasma-derived nutrients was inefficient . When an excess of exogenous methionine (200 microg/ml) was provided to the mtaR mutant, its growth rate in plasma was restored to that of the wild-type strain . The mtaR mutant grew poorly in chemically defined medium (CDM) prepared with methionine at a concentration similar to that of plasma (4 microg/ml) but was able to grow normally in CDM prepared with a high concentration of methionine (400 microg/ml) . Both the wild-type strain and the mtaR mutant were incapable of growth in CDM lacking methionine, indicating that GBS cannot synthesize methionine de novo . When the abilities of the strains to incorporate radiolabeled methionine were compared, the mtaR mutant incorporated fivefold less methionine than the wild-type strain during a 10-min period . Collectively, the results from this study suggest that the ability to regulate expression of a methionine transport system is critical for GBS survival in vivo. Blood, 2004 Mar 1, 103(5), 1702 - 9 Epub 2003 Oct 30. Thrombomodulin mutant mice with a strongly reduced capacity to generate activated protein C have an unaltered pulmonary immune response to respiratory pathogens and lipopolysaccharide; Rijneveld AW et al.; The thrombomodulin-protein C-protein S (TM-PC-PS) pathway exerts anticoagulant and anti-inflammatory effects . We investigated the role of TM in the pulmonary immune response in vivo by the use of mice with a mutation in the TM gene (TM(pro/pro)) that was earlier found to result in a minimal capacity for activated PC (APC) generation in the circulation . We here demonstrate that TM(pro/pro) mice also display a strongly reduced capacity to produce APC in the alveolar compartment upon intrapulmonary delivery of PC and thrombin . We monitored procoagulant and inflammatory changes in the lung during Gram-positive (Streptococcus pneumoniae) and Gram-negative (Klebsiella pneumoniae) pneumonia and after local administration of lipopolysaccharide (LPS) . Bacterial pneumonia was associated with fibrin(ogen) depositions in the lung that colocalized with inflammatory infiltrates . LPS also induced a rise in thrombin-antithrombin complexes in bronchoalveolar lavage fluid . These pulmonary procoagulant responses were unaltered in TM(pro/pro) mice, except for enhanced fibrin(ogen) deposition during pneumococcal pneumonia . In addition, TM(pro/pro) mice displayed unchanged antibacterial defense, neutrophil recruitment, and cytokine/chemokine levels . These data suggest that the capacity of TM to generate APC does not play a role of importance in the pulmonary response to respiratory pathogens or LPS. Vet Immunol Immunopathol, 2003 Dec 15, 96(3-4), 193 - 205 Dynamics of leukocytes and cytokines during experimentally induced Streptococcus uberis mastitis; Rambeaud M et al.; Streptococcus uberis causes a significant proportion of clinical and subclinical intramammary infections (IMI) in lactating and non-lactating dairy cows . In spite of this, its pathogenesis is incompletely understood . A study was conducted to determine leukocyte and cytokine dynamics during experimentally induced S . uberis mastitis . Five Jersey and five Holstein cows were challenged via intramammary inoculation of S . uberis into two uninfected mammary glands . Sixteen of 20 challenged mammary glands developed clinical mastitis with peak clinical signs observed at 144 h . The number of S . uberis in milk increased (P<0.05) 48 h after challenge, in spite of an increase in milk somatic cells that began at 18 h (P<0.001) and remained elevated throughout the study . Increased tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-8 (IL-8) in milk were detected 66 h after challenge (P<0.05) . Peak TNF-alpha and IL-8 concentrations occurred 120 h after challenge and preceded peak clinical signs . Experimental S . uberis IMI induced local production of TNF-alpha, IL-1beta and IL-8, which may play a role in the pathogenesis of S . uberis mastitis . Other mediators may be involved in initial leukocyte recruitment to the mammary gland, since increases in milk somatic cells occurred earlier than cytokine production. Arch Biochem Biophys, 2003 Nov 15, 419(2), 147 - 57 Analysis of recombinant acylated pneumococcal surface adhesin A of Streptococcus pneumoniae by mass spectrometry; De BK et al.; Streptococcus pneumoniae pneumococcal surface adhesin A (PsaA) is a species-common, immunogenic surface lipoprotein . In this study, the psaA gene was expressed as a nonfusion acylated protein in an Escherichia coli expression system . Yields of pure recombinant PsaA (rPsaA) were 8-10 mg/liter of fermentation culture . Analysis of rPsaA tryptic digests by HPLC-electrospray mass spectrometry (MS) confirmed 98% of the expected protein sequence . GC/MS data demonstrated very similar acylation of native and rPsaA by C12:0-C22:0 fatty acids, with C16 and C18 predominating . Negative ion electrospray MS/MS analysis of the rPsaA lipid anchor released by Pronase-E confirmed that the structure was based on an N-terminal palmitoylcysteine (Pam(3)Cys) . Electrospray MS heterogeneity analysis of intact rPsaA indicated that all of the observed heterogeneity could be accounted for by the fatty acid distributions . The availability of well-characterized rPsaA will facilitate the continued research and development of protein-based vaccines for the prevention of pneumococcal disease. Curr Biol, 2003 Oct 28, 13(21), 1858 - 66 The B cell SH2/PH domain-containing adaptor Bam32/DAPP1 is required for T cell-independent II antigen responses; Fournier E et al.; BACKGROUND: Bam32/DAPP1 is a B cell adaptor composed of both a PH and an SH2 domain . Previous studies in cell culture and chicken DT40 cells have indicated that Bam32 is critical for normal signaling downstream of the B cell receptor (BCR) . RESULTS: We now study the function of Bam32 in mice in which Bam32 has been disrupted by a viral gene trap approach . Although B and T cell development is normal in Bam32(-/-) mice, B cell proliferation is reduced by about 50% after BCR crosslinking when compared with Bam32(+/+) mice . Differences in the activation of Erk, Jnk and p38 Map kinases, PLCgamma, and Ca(2+) flux do not account for the defect in proliferation as activation was similar in Bam32(+/+) and Bam32(-/-) B cells . Interestingly, whereas antibody response to T-dependent (TD) and T-independent (TI)-I antigens was similar between Bam32(+/+) and Bam32(-/-) mice, TI-II responses were defective in Bam32(-/-) mice; Bam32(-/-) mice failed to undergo isotype class switch recombination (CSR) to produce IgG3 antibodies due to a cell-autonomous defect in generation of IgG3 germline transcripts . The defect in TI-II antigen response led to an impaired antibody response to immunization with type 3 Streptococcus pneumoniae capsular polyschaccharide (PS), resulting in a markedly increased susceptibility to infection by Streptococcus pneumoniae . CONCLUSIONS: These findings indicate that Bam32 specifically couples an upstream signal to the IgG3 isotype heavy chain CSR and suggest that defects in Bam32 may account for the increased susceptibility to encapusulated organisms in a subset of immunodeficient patients. J Coll Physicians Surg Pak, 2003 Oct, 13(10), 573 - 6 Mortality variables in pyogenic meningitis in paediatric age group; Khichi GQ et al.; OBJECTIVE: To determine the association of the studied variables on mortality in pyogenic meningitis in paediatric age group . DESIGN: An observational analytical study . PLACE AND DURATION OF STUDY: Paediatrics Medicine, Unit-2 of B.V . Hospital, attached with Quaid-e-Azam Medical College, Bahawalpur, from December 2000 to December 2002 . PATIENTS AND METHODS: Sample size was 300 admitted cases of pyogenic meningitis, aged 2 months to 12 years, meeting the inclusion criteria . The association of the different variables with mortality in pyogenic meningitis was studied . Data was collected and tabulated . X2 test of significance was applied to determine the p-value . p<0.05 was taken as statistically significant . RESULTS: The mortality rate was 10% (30/300) . The association of different variables, with outcome at the time of admission was statistically significant in prolonged duration of illness (chronicity prior to admission and increased cerebrospinal fluid (CSF) neutrophils after 1st lumbar puncture . Among the etiological microorganisms isolated from culture of CSF, only Streptococcus pneumoniae had statistically significant association . CONCLUSION: Mortality in pyogenic meningitis was significantly related to pre-admission illness duration and CSF neutrophilia . Mortality can be reduced by early and appropriate recognition . Antibiotic and supportive therapy should be initiated promptly followed by specific medication when the reports of the CSF culture is available. Surg Infect (Larchmt), 2003 Fall, 4(3), 255 - 62 In vitro antimicrobial activity of oxidized regenerated cellulose against antibiotic-resistant microorganisms; Spangler D et al.; BACKGROUND: The emergence of multi-drug resistant microorganisms presents a critical problem for patients undergoing surgery . Acidic pH, which is produced by oxidized regenerated cellulose (ORC), is a broad-spectrum physiological detriment to survival of microorganisms known to cause surgical infections . The purpose of this study was to examine the antimicrobial effect of ORC against antibiotic-resistant organisms . METHODS: ORC products were challenged with ATCC reference strains and clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE), vancomycin-resistant Enterococcus (VRE), penicillin-resistant Streptococcus pneumoniae (PRSP), and non-resistant ATCC strains of S . aureus and Pseudomonas aeruginosa . Samples of three ORC products (SURGICEL absorbable hemostat, SURGICEL Fibrillar absorbable hemostat, and SURGICEL NU-KNIT absorbable hemostat and identified, respectively, as ORC-R, ORC-F, and ORC-N for this study) were inoculated with challenge organisms in nutrient broth to produce a weight to volume ratio of 15 mg ORC/mL . Plate counts were performed at 0, 1, 6, and 24 h . RESULTS: Antimicrobial activity was seen with all three ORC products against the challenge organisms . Data indicate that antibiotic-resistant microorganisms remain susceptible to the antimicrobial activity of ORC . In testing with nine of 10 bacteria, including four antibiotic-resistant clinical isolates (VRE, MRSA, and PRSP) three-log or greater reductions were seen at 24-h exposure . One ATCC strain of VRE demonstrated some level of resistance to the acidic pH effect . ORC-N showed a three-log reduction with this organism, whereas the reductions with ORC-R and ORC-F were less than one log . CONCLUSION: Since low pH affects a relatively broad-spectrum of bacteria and does not act in a mechanism-specific manner, as do antibiotics, antibiotic-resistant strains of bacteria are unlikely to resist the ORC pH effect . Results of this in vitro assessment support the hypothesis that the antimicrobial activity of ORC is effective against antibiotic-resistant microorganisms. J Oral Maxillofac Surg, 2003 Oct, 61(10), 1156 - 61 The presence of bacteria in the synovial fluid of the temporomandibular joint and clinical significance: preliminary study; Kim SJ et al.; PURPOSE: The objective of this study was to find any relation between the presence of specific bacterial species in the synovial fluid of the temporomandibular joint (TMJ) and clinical parameters . PATIENTS AND METHODS: We studied 43 patients (male-to-female ratio, 1:1.69; average age, 34.37 +/- 14.55 years) . Thirty-three patients had a displaced disc in the TMJ (DD group), and 10 patients did not have a displaced disc of the TMJ or any symptom related to TMJ disorders (NDD group) . Clinical examinations were made to determine maximum mouth opening, joint sounds, previous trauma history, systemic disease, and TMJ pain . Six bacterial species that were reported in other studies were chosen to evaluate the presence of bacteria in the TMJ for this study . RESULTS: Mycoplasma genitalium was most frequently detected in synovial fluid (86.0%) . Staphylococcus aureus, Mycoplasma fermentans/orale, Actinobacillus actinomycetemcomitans, and Streptococcus mitis were detected in 51.2%, 37.2%, 25.6%, and 7.0% of samples, respectively . beta-Hemolytic Streptococcus was not detected . The prevalence of S aureus was significantly higher in the DD group than in the NDD group (P <.05) . The patients who had M . fermentans/orale were 5.40 times more likely to be younger than 30 years than were those without M . fermentans/orale (P <.05) . Those with M . genitalium were 5.81 times more likely to be female than were those without M . genitalium (P <.05) . CONCLUSION: The presence of S . aureus in TMJ synovial fluid was related to TMJ disorder symptoms and clinical parameters seemed to be influenced by bacterial presence in TMJ synovial fluid. J Antimicrob Chemother, 2003 Dec, 52(6), 944 - 52 Epub 2003 Oct 29. Worldwide incidence, molecular epidemiology and mutations implicated in fluoroquinolone-resistant Streptococcus pneumoniae: data from the global PROTEKT surveillance programme; Canton R et al.; OBJECTIVES: To analyse the mutations and epidemiology associated with fluoroquinolone-resistant pneumococci collected as part of the PROTEKT global surveillance programme during 1999-2000 . METHODS: Sixty-nine centres in 25 countries submitted a total of 3362 Streptococcus pneumoniae isolates, for which the MICs of antimicrobial agents were determined using NCCLS methodology . RESULTS: Levofloxacin resistance was low overall (1% worldwide), with higher rates in: Hong Kong (14.3%), South Korea (2.9%), USA (1.8%), Mexico (1.5%), Canada (1.4%) and Japan (1.3%) . Levofloxacin resistance was very low or absent in European countries, and absent in Australia . Worldwide, there was a total of 35 levofloxacin-resistant isolates, of which 22 (63%) were resistant and 10 (29%) were intermediate to moxifloxacin . All levofloxacin-resistant isolates were susceptible to telithromycin (< or =0.5 mg/L), linezolid (< or =2 mg/L) and quinupristin/dalfopristin (< or =1 mg/L) . One or more mutations in the topoisomerase genes were identified in all levofloxacin-resistant isolates; most of these isolates (33/35) had a mutation in one of the DNA gyrase encoding genes (gyrA, gyrB) and one of the topoisomerase IV encoding genes (parC, parE) . Eighteen (51%) isolates carried the same combination of amino acid substitutions: Ser-81-->Phe in GyrA and Ser-79-->Phe in ParC . Isolates displaying a levofloxacin MIC of 2-4 mg/L generally had no mutation or one mutation in either a DNA gyrase or a topoisomerase IV gene, although most mutations were in parC . CONCLUSIONS: Most levofloxacin-resistant isolates possess two mutations (one in DNA gyrase and one in topoisomerase IV) . Although multilocus sequence typing demonstrated that most of these isolates were unrelated, 12 (34%) were the Spain23F-1 clone: 10 from Hong Kong and one each from Saskatchewan, Canada and Sao Paulo, Brazil. J Antimicrob Chemother, 2003 Dec, 52(6), 953 - 60 Epub 2003 Oct 29. Emergence of a fluoroquinolone-resistant strain of Streptococcus pneumoniae in England; Johnson AP et al.; OBJECTIVE: To determine the epidemiological relationship between pneumococci of serotype 9V, with reduced susceptibility to ciprofloxacin, penicillin and erythromycin, referred to the Reference Laboratory during 1997-2001 . METHODS: Isolates were characterized by multilocus sequence typing (MLST), PFGE, and sequencing of parC and gyrA . Relevant clinical data were sought . RESULTS: Forty-eight isolates were received from nine laboratories in England, but 35 (73%) were from one laboratory in Birmingham, and were mostly from elderly patients receiving ofloxacin or ciprofloxacin for respiratory infections . There were two quinolone resistance phenotypes, with ciprofloxacin, moxifloxacin and gemifloxacin MICs of 8-32, 0.5-1 and 0.125-0.25 mg/L, and 64-256, 4-16 and 1-4 mg/L, respectively . Each of three isolates from the former group had mutations in parC, whereas each of nine isolates from the more resistant group had mutations in both parC and gyrA . Several also had increased quinolone efflux . Typing of 27 quinolone-resistant isolates showed that eight were indistinguishable from the epidemic Spain9V-3 (ST156) clone, while the remainder belonged to a novel but related type (ST609), that differed from Spain9V-3 at 2/7 alleles (2 bp changes in aroE and 1 bp change in gdh) . Both MLST types were represented among isolates with high- and low-level quinolone resistance . Three of five serotype 9V isolates from Birmingham, with reduced susceptibility to penicillin and erythromycin, and ciprofloxacin MICs of 1-2 mg/L, belonged to MLST type ST609, while another was indistinguishable from the Spain9V-3 clone . Review of records of 32 patients from Birmingham indicated that some isolates were nosocomial, whereas others were acquired in the community . CONCLUSIONS: In the late 1990s, a quinolone-resistant strain, clonally related to Spain9V-3, emerged in England, principally in Birmingham. Cochrane Database Syst Rev . 2003;(4):CD000422. Vaccines for preventing pneumococcal infection in adults; Dear K et al.; BACKGROUND: Diseases caused by Streptococcus pneumoniae (S . pneumoniae) continue to cause substantial morbidity and mortality throughout the world . Polysaccharide pneumococcal vaccines have been developed for over 50 years and may have the potential to prevent disease and death . OBJECTIVES: To assess the effectiveness of polysaccharide pneumococcal vaccination in preventing disease or death in adults . SEARCH STRATEGY: Trials were identified by electronic searches of the Cochrane Central Register of Controlled Trials (CENTRAL) issue 2, 2003 (which includes the Cochrane ARI Group's specialised register); MEDLINE (January 1966 to June 2003); and EMBASE (1974 to June 2003) . We searched existing literature . The bibliographies of all newly revealed studies were read in order to identify further studies . The vaccine manufacturers, the lead authors of newly identified studies not included in existing meta-analyses were contacted . SELECTION CRITERIA: A) Prospective, randomised or quasi-randomised studies comparing pneumococcal vaccines with placebo, control vaccines or no intervention.B) Case-control studies (including indirect cohort studies) assessing pneumococcal vaccine effectiveness against invasive pneumococcal disease . Cohort studies are excluded . DATA COLLECTION AND ANALYSIS: A) Randomised studies . Trial quality assessment was conducted by two reviewers (JH and DT) . Data extraction was done by three reviewers (JH, DT, KD) . There were many instances of unclear or incomplete data in the trial reports, and the final dataset was arrived at after much deliberation and discussion, including comparison with the data used in two previous reviews of this question . Due to the age of the trials (dating back to 1954 in one case) it was not generally possible to obtain clarification from the authors, though a partial clarification was achieved in one case.B) Non-randomised studies . Study quality was assessed by two reviewers (RA and KD) . MAIN RESULTS: The combined results from the randomised studies fail to show that the polysaccharide pneumococcal vaccine is effective in preventing either pneumonia (outcome 6: odds ratio = 0.77, confidence interval 0.58, 1.02, number = 14) or death (outcome 8: odds ratio 0.90, confidence interval 0.76, 1.07, number = 11) . Despite encouraging data from some very early trials, pooling trials published from 1977 on suggests there is no effect (outcome 6; odds ratio = 0.96, confidence interval 0.80, 1.15, number = 12; outcome 9: odds ratio = 0.98, confidence interval 0.88, 1.09, number = 10) . The available data cannot distinguish whether this heterogeneity in results is due to improvements in trial methodology and reporting, to differences in trial setting or to real loss of efficacy over time . This is because the early, poorly reported trials were conducted in high-risk healthy populations where the expected benefit is greatest.The case-control studies show significant efficacy in preventing invasive pneumococcal disease: OR 0.47 (CI 0.37, 0.59) corresponding to an efficacy of 53% . REVIEWER'S CONCLUSIONS: While polysaccharide pneumococcal vaccines do not appear to reduce the incidence of pneumonia or death in adults with or without chronic illness, or in the elderly (55 years and above), the evidence from non-randomised studies suggests that the vaccines are effective in the reducing the incidence of the more specific outcome, invasive pneumococcal disease, among adults and the immunocompetent elderly (55 years and above) . Surveillance data suggest that infection rates vary widely between and also within countries, but a typical figure in developed countries is 0.01%, or 10 per 100,000 per year . Efficacy of 50% then corresponds to a number-needed-to-treat (NNT) of 20,000 vaccinations per infection avoided, and perhaps 50,000 per death avoided. Clin Infect Dis, 2003 Nov 15, 37(10), 1335 - 9 Epub 2003 Oct 13. Pneumonia in older persons; Loeb M; Community-acquired pneumonia (CAP) is an important threat to the health of older adults . Streptococcus pneumonia remains the most important cause of CAP . Risk factors for CAP include alcoholism, asthma, immunosuppression, chronic respiratory or cardiac disease, institutionalization, and increasing age . Residents of long-term care facilities--a distinct subpopulation of elderly people--are at particularly high risk for developing pneumonia . In this setting, swallowing difficulties, witnessed aspiration, and receipt of sedatives are potentially modifiable risk factors . The clinical presentation in elderly patients is characterized by a reduced prevalence of nonrespiratory symptoms . Few randomized, controlled trials of therapy exist for elderly persons living in the community or in a long-term care setting . Good evidence exists to support the annual administration of influenza vaccine to older adults . Although evidence in clinical trials differs from evidence in observational studies that demonstrate clear benefits associated with the polysaccharide pneumococcal vaccine in this population, the vaccine is recommended for adults aged J Zoo Wildl Med, 2003 Sep, 34(3), 296 - 301 A third report of "golf ball disease" in an Amazon River dolphin (Inia geoffrensis) associated with Streptococcus iniae; Bonar CJ et al.; An Amazon River dolphin (Inia geoffrensis) developed a dermatologic syndrome characterized by the occurrence of slow-growing, nodular, s.c . abscesses . Initial biopsies, cultures, and cytologic analysis of needle aspirates from the abscesses indicated steatitis with probable secondary, gram-negative bacterial infection . Treatment with dietary vitamin E supplement and broad-spectrum antibiotics yielded minimal improvement . Subsequent cultures revealed Streptococcus iniae in addition to several gram-negative bacteria . Vigorous surgical management of the abscesses, including lancing, debridement, and irrigation, combined with antimicrobial therapy specific for Streptococcus and gram-negative organisms, and improvement of the animal's diet and environmental water quality led to gradual recovery . When the animal was ill, it demonstrated an inflammatory leukogram and transient uremia . Streptococcus iniae is a serious pathogen of aquacultured fishes and humans and should be included in the differential diagnosis of chronic dermatopathy in river dolphins . Specific antimicrobial therapy, excellent water quality, surgical management of abscesses, and adherence to sanitary protocols should be observed in cases of suspected S . iniae infection in dolphins. Scand J Plast Reconstr Surg Hand Surg, 2003, 37(4), 239 - 42 Necrotising fasciitis and myositis that originated from gastrointestinal bacterial infection: two fatal cases; Fujioka M et al.; We present two cases of aggressive myofasciitis after gastrointestinal infections . The first patient developed necrosis of the legs over 5 days and he died on the 20th day . Vibrio vulnificus was cultured . The second patient developed bullae on his thigh and he died the next day . Streptococcus pyogenes was cultured. Acta Odontol Scand, 2003 Aug, 61(4), 241 - 6 Antimicrobial effects of glass ionomer cements containing bioactive glass (S53P4) on oral micro-organisms in vitro; Yli-Urpo H et al.; The antimicrobial effects of two glass-ionomer cements (GICs), GC Fuji II and Fuji II LC, mixed with different amounts (0 wt%, 10 wt% and 30 wt% of the total powder weight) of bioactive glass (BAG), S53P4, on Streptococcus mutans and Candida albicans were studied in vitro . The growth inhibition was tested using agar diffusion . The materials were also studied in a liquid media . The effect of the material extracts on acid production was studied using cell suspensions . The antimicrobial activity of the materials was examined by incubating the cell suspensions with the material powder . In the agar diffusion test, only the GICs containing 30 wt% BAG inhibited the growth of S . mutans . When the materials were tested in culture medium, no inhibitory effects on S . mutans were detected . The only materials to inhibit acid production of S . mutans were the GIC extracts without added BAG . Furthermore, they also had antibacterial activity against S . mutans when tested as powders . We found very few effects of the tested materials on C . albicans . The only material with an antimicrobial effect on C . albicans was BAG when incubated in a suspension with C . albicans . This is the first time that this effect has been demonstrated for C . albicans . By adding BAG to GICs the structure of the material becomes more brittle than the structure of GICs without BAG . Thus, in addition to the composition of the tested materials, their structure may also have influenced the results . In summary, commercially available GICs and GIC disks containing 30 wt% of BAG exerted antibacterial effects on S . mutans . BAG exerted antimicrobial effects on both S . mutans and C . albicans. J Microbiol Immunol Infect, 2003 Sep, 36(3), 175 - 81 Rate of nasopharyngeal carriage, antimicrobial resistance and serotype of Streptococcus pneumoniae among children in northern Taiwan; Lo WT et al.; Multiple-antibiotic-resistant strains of Streptococcus pneumoniae are isolated from clinical specimens in Taiwan with increasing frequency . This study aimed to define the carriage rate of S . pneumoniae among children in northern Taiwan, and to determine the antibiotic susceptibility and the serotype incidence of these isolates . Nasopharyngeal swabs were taken from a total of 478 children (age, 1 month-14 years) who sought medical care only for non-infectious disease or routine vaccination at our hospital between July 1998 and November 1999 . S . pneumoniae was isolated from 95 patients, and the collected isolates were available for analysis . All pneumococcal isolates were serotyped and their antimicrobial susceptibility tested by standard methods . The total rate of pneumococcal carriage in the study population was 19.9% and the isolation rate was higher in children aged between 2 and 5 years . Only 10 (10.5%) of the isolates were susceptible to penicillin (minimum inhibitory concentration {MIC}, < or = 0.06 microg/mL); 47 (49.5%) isolates were intermediately resistant (MIC, 0.12-1 microg/mL) and 38 (40%) were highly resistant (MIC, > or = 2 microg/mL) . Among the 95 S . pneumoniae isolates, the common serotypes were 23F (22%), 6B (18.9%), 19F (18.9%), and 14 (8.4%) . Evaluation of the results showed that serotypes 23F (24.7%), 19F (21.2%), 6B (15.3%), and 14 (9.4%) composed 70.6% of all penicillin-non-susceptible S . pneumoniae isolates . The significant rate of isolation of penicillin-non-susceptible S . pneumoniae from children indicates that both the judicious use of antibiotics and the availability of conjugate pneumococcal vaccines are the most appropriate strategy to reduce the carriage of resistant pneumococci. Indian Pediatr, 2003 Oct, 40(10), 1002 - 4 Varicella complicated by scarlet fever; Yavuz T et al.; We report a 3-year-old boy with varicella complicated by cellulitis and scarlet fever . He developed a typical rash of scarlet fever following the onset of varicella . Streptococcus pyogenes was isolated from the ulcers due to varicella . The present case suggests that scarlet fever may rarely develop following varicella and should be considered in children with complicated varicella. Int J Food Microbiol, 2003 Dec 15, 89(1), 77 - 83 Inhibition of Listeria monocytogenes by bovicin HC5, a bacteriocin produced by Streptococcus bovis HC5; Mantovani HC et al.; Cattle can be infected with Listeria monocytogenes by consuming contaminated plant materials, soil or silage, and farmers have sought ways of preventing this contamination . Recent work indicated that Streptococcus bovis HC5 produced a bacteriocin (bovicin HC5) that could inhibit a variety of gram-positive bacteria, and we examined the ability of bovicin HC5 to inhibit 10 strains of L . monocytogenes that had been isolated from plant materials, soil, silage and infected cattle . Growth experiments indicated that all of the L . monocytogenes strains were inhibited by 100 activity units (AU) of bovicin HC5 ml(-1) . L . monocytogenes cultures that were transferred with sublethal doses (12.5 AU ml(-1)) could be adapted in stepwise fashion to higher doses of bovicin HC5 . However, even 'adapted' cultures did not grow if 400 AU ml(-1) was added . The effect of bovicin HC5 on L . monocytogenes was bactericidal, and viability decreased 5-7 logs after only 2 h of exposure . Bovicin HC5 caused a nearly complete efflux of intracellular potassium in 15 min but only if the pH was less than 6.0 . When the pH was greater than 6.0, the cells maintained their potassium pool . L . monocytogenes cells that were acid-adapted (final pH of 4.6) were as sensitive to bovicin HC5 as those that were not acid-adapted (final pH of 6.3) . These results support the idea that bovicin HC5 could be effective in controlling listeria in contaminated silages. Clin Chem, 2003 Nov, 49(11), 1839 - 45 Apolipoprotein C-III isofocusing in the diagnosis of genetic defects in O-glycan biosynthesis; Wopereis S et al.; BACKGROUND: Defects in the biosynthesis of N-glycans may be found by isoelectric focusing (IEF) of plasma transferrin . No test is available to demonstrate O-glycan biosynthesis defects . METHODS: We used isoforms of apolipoprotein C-III (apoC-III) as a marker for the biosynthesis of core 1 mucin type O-glycans . Plasma samples from patients with primary defects and secondary alterations in N-glycan biosynthesis were studied by apoC-III isofocusing . RESULTS: Age-related reference values for apoC-III were determined . Plasma samples from patients with the primary congenital disorders of glycosylation (CDG) types Ia-Ic, Ie, If, IIa, and IId all showed a normal apoC-III isofocusing profile . Plasma from two patients with CDG type IIx were tested: one showed a normal apoC-III distribution, whereas the other showed a hypoglycosylation profile . In plasma from patients with hemolytic uremic syndrome (HUS), a hypoglycosylation profile was obtained . CONCLUSIONS: IEF of apoC-III is a rapid and simple technique that may be used as a screening assay for abnormalities in core